bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2936: JNJ-47117096, a selective small molecule inhibitor of the MELK oncogene decreases DNA damage tolerance in highly proliferating cancer cells
Authors: Beke Lijs , Linders Joannes T.m. , Kig Cenk , Boeckx An , Heerde Erika van , Wuyts Dirk , Parade Marc , Meerpoel Lieven , Johnson Chris , Beullens Monique , Bollen Mathieu , Brehmer Dirk
Published: May 2015

Maternal embryonic leucine zipper kinase (MELK, hMP38, pEG3), a Ser/Thr protein kinase, is highly overexpressed in stem cells and cancer cells. The oncogenic role of MELK is attributed to disabling critical cell cycle check points as well as enhancing replication. Most functional studies have relied on the use of siRNA/shRNA mediated gene silencing, but this often can be associated with off-target effects.

Here we want to present a novel, potent and selective small molecule inhibitor JNJ-47117096 that has enabled us to validate the biological function of MELK kinase.

Outcome: MELK inhibition in cancer cells with an intact p53 signaling pathway is linked to a replicative senescence phenotype. This phenotype correlates with a rapid ATM activation and phosphorylation of CHK2 without any effects on the alternative ATR/ CHK1 DNA damage pathways. Furthermore, JNJ-47117096 induces strong phosphorylation of p53 and prolonged up-regulation of p21 without the induction of apoptosis. Strikingly, MELK inhibition in several p53 disabled cancer cells showed induction of a mitotic catastrophe phenotype followed by prominent cell killing.

Finally, JNJ-47117096 triggers a rapid degradation of cellular MELK protein, independent of the cell cycle phase and its regulation by the E2F pathway. This observation can clearly be linked to a direct binding effect of JNJ-47117096 to cellular MELK protein confirmed by chemical proteomics.

Conclusion: Our data generated with JNJ-47117096, confirmed by selective siRNAs, indicate MELK as a key stimulator of proliferation and replication by its ability to increase the threshold for DNA damage tolerance. Targeting MELK function by selective small molecule inhibitors might sensitise tumors to DNA-damaging agents or radiation therapy.

Citation Format: Lijs Beke, Joannes T.m. Linders, Cenk Kig, An Boeckx, Erika van Heerde, Dirk Wuyts, Marc Parade, Lieven Meerpoel, Chris Johnson, Monique Beullens, Mathieu Bollen, Dirk Brehmer. JNJ-47117096, a selective small molecule inhibitor of the MELK oncogene decreases DNA damage tolerance in highly proliferating cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2936. doi:10.1158/1538-7445.AM2014-2936
Abstract 3020: Patient-derived xenograft (PDX) models to study the role of breast cancer stem cells in metastasis formation
Authors: Ginestier Christophe , Lartigue Danae , Cabaud Olivier , Malzac Aurelie , Wicinski Julien , Josselin Emmanuelle , Finetti Pascal , Adelaide Jose , Bertucci Francois , Chaffanet Max , Birnbaum Daniel , Charafe-Jauffret Emmanuelle
Published: May 2015

Although advances have been made in the treatment of localized breast cancer, there has been less progress in the treatment of advanced metastatic disease. This disease is an oncoming therapeutic challenge for breast cancer since it is always fatal. Conventional therapies may fail to target cancer stem cells (CSC). Metastatic breast cancer may thus benefit from current intensive research in the CSC field. Several studies suggest that CSCs are the only cancer cells responsible for metastasis formation. CSCs may have the ability to disseminate in blood circulation as circulating tumor cells (CTCs) and to colonize new sites. To better understand the biological processes that link CSCs and metastasis, we have generated a xenobank of 50 patient-derived xenografts (PDXs) of primary tumors grafted in the humanized mammary fat pad of immunodeficient mice. These models reflect the heterogeneity of primary tumors both at molecular and cellular level. Using a lentiviral vector, we infected our PDXs to obtain GFP-luciferase-positive models designed to monitor metastasis development from the orthotopic site. To mimick the patient therapeutic management, metastasis formation is monitored during tumor growth but also after tumor surgical removal. Metastases are collected and analyzed for histology and phenotype. The different metastatic patterns are correlated with the mutational profile of PDX. CTCs are also monitored at different time of the metastatic process using a specific filter (ISET) for cell characterization and phenotype. PDXs can be classified in tumor groups with similar metastatic tropisms. To depict the role of CSC from primary tumors in the tropism of their metastases, we collected and genomic profiled the ALDEFLUOR-positive CSCs in orthotopic tumors from different models with distinct metastatic pattern. These different PDX models lead to a better understanding of the role of breast CSCs in metastasis formation from CTCs to well-constituted metastasis and their study may identify targets able to design future therapeutic strategies.

Citation Format: Christophe Ginestier, Danae Lartigue, Olivier Cabaud, Aurelie Malzac, Julien Wicinski, Emmanuelle Josselin, Pascal Finetti, Jose Adelaide, Francois Bertucci, Max Chaffanet, Daniel Birnbaum, Emmanuelle Charafe-Jauffret. Patient-derived xenograft (PDX) models to study the role of breast cancer stem cells in metastasis formation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3020. doi:10.1158/1538-7445.AM2014-3020
Abstract 3035: Cripto/GRP78 signaling promotes the stem cell phenotype in normal and neoplastic mammary epithelial cells
Authors: Spike Benjamin T. , Kelber Jonathan A. , Booker Evan , Kalathur Madhuri , Rodewald Rose , Lipianskaya Julia , La Justin , He Marielle , Wright Tracy , Klemke Richard , Wahl Geoffrey , Gray Peter C.
Published: May 2015

Microenvironmental factors are critical regulators of stem cell and tumor cell behavior. However, few stem cell factors have been identified and targeted in breast and other cancers. Using a highly enriched stem cell pool isolated from fetal mammary rudiments and single cell analyses, we have identified the onco-fetal protein Cripto as a potent soluble factor that regulates the mammary stem cell state. Cripto is a GPI-anchored/secreted signaling protein and a known regulator of PI3K/AKT and TGF-beta pathways. Here, we develop a novel Cripto antagonist, ALK4L75A-Fc, which selectively blocks the growth factor-like effects of soluble Cripto. We show that this antagonist promotes the differentiation of mammary stem cells cultured ex vivo while Cripto treatment maintains the stem cell phenotype and yields colonies with enhanced mammary gland reconstitution capacity. We previously discovered that Cripto signaling requires binding to cell surface Glucose Response Protein 78kDa (GRP78). GRP78 is an HSP70 family member that is induced by stresses including hypoxia and glucose deprivation and it is highly expressed in tumors where these conditions prevail. We show here that cell surface GRP78 marks fetal mouse mammary stem cells and a population of bipotent adult mammary epithelial cells that are selectively responsive to soluble Cripto in vitro. GRP78high mammary epithelial cells also harbor substantially higher stem cell activity upon transplantation than GRP78low cells. Consistently, we further show that deletion of GRP78 from adult mammary epithelial cells ex vivo blocks mammary gland reconstitution. Finally, we find that Cripto antagonism with ALK4L75A-Fc inhibits the proliferation of triple negative breast cancer cell lines in vitro and that this antagonist can also inhibit tumor growth in vivo. Since Cripto and GRP78 are both induced by hypoxia and other stressful conditions found in tumors, our data raise the possibility that Cripto/GRP78 signaling may exacerbate breast and other cancers by increasing the number of stem cell-like tumor cells in tumor microenvironments where these conditions predominate.

Citation Format: Benjamin T. Spike, Jonathan A. Kelber, Evan Booker, Madhuri Kalathur, Rose Rodewald, Julia Lipianskaya, Justin La, Marielle He, Tracy Wright, Richard Klemke, Geoffrey Wahl, Peter C. Gray. Cripto/GRP78 signaling promotes the stem cell phenotype in normal and neoplastic mammary epithelial cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3035. doi:10.1158/1538-7445.AM2014-3035
Abstract 3046: Lin28B mediated IKK-β sustains the stemness of breast cancer stem cell via regulating Wnt/TCF4 and miR-34a/LEF1 signaling pathway
Authors: Chen Chong , Bai Lipeng , Cao Fengqi , Liu Yan , Xie Junling , Wang Wei , Si Qin , Yang Jian , Chang Antao , Xiang Rong , Luo Yunping
Published: May 2015

Substantial evidence indicates that many human cancers are driven by a subpopulation of cells that display stem cell properties. These cancer stem cells (CSCs) may also contribute to tumor metastasis and treatment resistance. It has been reported that inflammatory factors in tumor microenvironment activate NF-kB/IKK-β pathway in tumor cells which in turn drive CSC self-renewal, but their molecular basis remain unclear. In this study, we demonstrated a novel mechanism regarding that Lin28B, a RNA binding protein regulated by IKK-β, sustains the stemness of breast cancer stem cell via regulating Wnt/TCF4 and miR-34a/LEF1 signaling pathway. First, we found that all stemness properties of MDA-MB-231 human breast cancer cells including ALDH+ population, Sphere-forming ability and the expression of stemness genes SOX2, OCT4, NANOG and Lin28B were decreased by IKK-β inhibition, meanwhile, the down regulated expression of Lin28B decrease the stemness properties of MDA-MB-231 cells as well. Furthermore, it was found that Wnt/TCF4 signaling pathway was inhibited by Lin28B down-regulation via reducing the expression of TCF4. Interestingly, TCF4 could also regulate the expression of Lin28B by directly binding to the intron 1 of Lin28B. Moreover, we observed that microRNA-34a was down regulated by Lin28B at transcriptionally level, and increased miR34a expression inhibited the activity of Wnt/LEF1 pathway by directly targeting LEF1. Most importantly, we found that down regulated expression of Lin28B or IKK-β inhibition combined with chemo-drug (IKK-β selective inhibitor IMD-0354 combine with paclitaxel) could markedly suppressed tumor growth and metastasis of MDA-MB-231 breast tumor cells in NOD/SCID mice model. Taken together, our study demonstrated that Lin28B, which was activated by IKK-β, could sustain the stemness of CSC via the TCF4 and miR-34a/LEF1 signaling regulation, thus, establish the Lin28B as a central regulator in govern inflammation to maintain breast cancer stemness phenotype, and could serve as a therapeutical target.

Citation Format: Chong Chen, Lipeng Bai, Fengqi Cao, Yan Liu, Junling Xie, Wei Wang, Qin Si, Jian Yang, Antao Chang, Rong Xiang, Yunping Luo. Lin28B mediated IKK-β sustains the stemness of breast cancer stem cell via regulating Wnt/TCF4 and miR-34a/LEF1 signaling pathway. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3046. doi:10.1158/1538-7445.AM2014-3046
Abstract 3055: Potential role of cyclin D1 in regulation of liver cancer stem cells
Authors: Xia Wei , Wang Xiao Qi
Published: May 2015

Cancer stem cells (CSCs) are capable of self-renewal, multipotency, in vivo tumorigenicity, and driving metastasis. CSCs are more resistant to conventional therapy, allowing recurrence of tumor. Thus, novel therapies capable of eliminating CSCs are needed. Cyclin D1 is deregulated in many types of cancers. Beyond being a cell cycle regulator and oncogene, cyclin D1 also enhances stem cell (SC) self-renewal and promotes induced pluripotent stem cells (iPSCs) reprogramming efficiency. Yet, the role of cyclin D1 in regulation of CSC is not well defined.

Here we investigated whether and how cyclin D1 could regulate liver cancer CSC properties. We overexpressed cyclin D1 in two lines of HCC cells, Huh7 and 97H and found that the sphere formation capacity was enriched in Huh7 cyclin D1-expressing cells than Huh7 cells. From dissected single spherical cells, the secondary sphere formation capacity was significantly higher in cyclin D1-expressing cells than both parental Huh7 (p=0.00) and 97H (p=0.02) cells. Anchorage-independent spherical colonies (spheres) are composed of a small number of cancer cells with stem-like properties. Thus, the results suggest that overexpression of cyclin D1 enhanced self-renewal capacity of liver CSCs. The cyclin D1-expressing spheres displayed an increased expression of stem cell-associated genes Nanog, Oct4, and Nordal/Activin. Smad2/3, a TGF-β signaling effector, was highly phosphorylated in cyclin D1-expressing spheres compared to monolayer or spheres without cyclin D1 expression.

More importantly, TGF-β/Smad inhibitor (SB431542) inhibited cyclin D1-expressing CSC self-renewal shown by significantly reduction of sphere formation in two lines of cyclin D1-expressing HCC lines, whereas it had no effects on sphere formation in parental cells without cyclin D1 overexpression. In addition, Erk inhibitor (UO126) showed no inhibiting effects, a further proof of the role of cyclin D1 mediated activation of Smad2/3. These results indicate that TGF-β/Smad inhibitor might have therapeutic potential for targeting liver CSCs.

HCC cells are highly resistance chemotherapy resulting in enriched CSC population. We further investigated the sphere formation capacity of D1-expressing cells under the treatment of chemotherapeutic agents in combination with Smad inhibitor. We found that SB431542 could inhibit cyclin-D1 expressing spherical cancer cell proliferation therefore sensitized the cells to chemo drug cisplatin. In summary, our results suggest that cyclin D1 might promote liver CSC proliferation and self-renewal via activation of TGF-β/Smad signaling, with underlining mechanism is under investigation.

Citation Format: Wei Xia, Xiao Qi Wang. Potential role of cyclin D1 in regulation of liver cancer stem cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3055. doi:10.1158/1538-7445.AM2014-3055
Abstract 3056: Human ovarian cancer stem cells: In vitro cultivation and characterization
Authors: Ishiguro Tatsuya , Ohata Hirokazu , Nakagama Hitoshi , Okamoto Koji , Tanaka Kenichi , Enomoto Takayuki
Published: May 2015

Cancer stem cells (CSC) constitute a miniscule population within the heterogeneous cells of cancer tissues, but they can self-renew, differentiate and recapitulate the tumors. In the absence of serum, CSC from some solid tumors are capable of proliferation in vitro under sphere-forming conditions. We have now established spheroid culture conditions for cultivating ovarian cancer cells derived from surgical specimen. These spheroid cells are tumorigenic when explanted into NOD/Shi-scid, IL-2Rγnull mice, and immuno-histological analysis indicates that the resulting tumors are indistinguishable from the originals. When serum is added to the in vitro culture media, the spheroid cells differentiate, which is associated with the down-regulation of the stem cell-specific regulators Nanog, Sox2, and ALDH1A1 and the up-regulation of cytokeratin. Expression of two of the regulators was essential for the sphere cell maintenance because shRNA-mediated knockdown of either Sox2 or ALDH1A1 inhibited cell proliferation. The differentiated cells, when returned to the sphere-forming serum-free conditions, were capable of reforming spheroids and re-expression of the stem cell markers. Single cell gene expression analysis of spheroid cells and the differentiated cells revealed that these two cell populations were clearly different, and that the reverting spheroid cells were similar to the original spheroid population. These findings indicated that the differentiation of the spheroid stem cells was highly reversible. Unexpectedly, the serum-induced differentiation caused the cell population to become more resistant to paclitaxel, one of the key ovarian cancer chemotherapy drugs, whereas de-differentiation/reversion back to spheroids rendered the cells paclitaxel-sensitive again. The intriguing clinical implications of these findings will be discussed.

Citation Format: Tatsuya Ishiguro, Hirokazu Ohata, Hitoshi Nakagama, Koji Okamoto, Kenichi Tanaka, Takayuki Enomoto. Human ovarian cancer stem cells: In vitro cultivation and characterization. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3056. doi:10.1158/1538-7445.AM2014-3056
Abstract 3075: Ex-vivo expansion of circulating tumor cells in early lung cancer using a microfluidic model
Authors: Zhang Jennifer Zhuo , Shiratsuchi Hiroe , Lin Jules , Chen Guoan , Reddy Rishindra M. , Azizi Ebrahim , Fouladdel Shamileh , Chang Andrew C. , Lin Lin , Simeone Diane M. , Wicha Max S. , Beer David G. , Ramnath Nithya , Nagrath Sunitha
Published: May 2015

Circulating tumor cells (CTCs) have gathered much momentum in recent times with emerging micro and nanotechnologies enabling better recoveries of these rare cells. Future research directions in CTCs significantly depend on obtaining meaningful number of CTCs that are available for downstream assays. This can be achieved by expanding CTCs. However, in early stage cancer patients, the numbers of cells that circulate in the peripheral blood are rare. Therefore until now, expanding CTCs remains to be a serious challenge limiting clinical utility. Microfluidic technologies have demonstrated that these rare cells can be recovered with better efficiency, purity and high viability. Herein, we developed a strategy using microfluidic technology to isolate and expand CTCs. We have successfully expanded CTCs isolated from 16 out of 21 early stage lung cancer patients. Expanded CTCs expressed tumor specific markers and formed spheroids in culture. Furthermore, these expanded CTCs carried the identical cancer related mutations (e.g. TP53 mutation) as their primary tumors. We expect that our model opens up the unprecedented avenues to understand biology of these lethal drivers of metastasis offering opportunities for personalized medicine through target drug evaluations.

Citation Format: Jennifer Zhuo Zhang, Hiroe Shiratsuchi, Jules Lin, Guoan Chen, Rishindra M. Reddy, Ebrahim Azizi, Shamileh Fouladdel, Andrew C. Chang, Lin Lin, Diane M. Simeone, Max S. Wicha, David G. Beer, Nithya Ramnath, Sunitha Nagrath. Ex-vivo expansion of circulating tumor cells in early lung cancer using a microfluidic model. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3075. doi:10.1158/1538-7445.AM2014-3075
Abstract 3985: Differential activation of unfolded protein response in metastatic vs. non-metastatic osteosarcoma cells following hypoxia and chemotherapeutic stress
Authors: Faraj Janine , Hingorani Pooja , Sertil Aparna R.
Published: May 2015

Osteosarcoma (OS) is the most common primary tumor of bone and the second highest cause of cancer-related death in the pediatric age group. Despite significant advances in diagnosis and treatment for this cancer, the overall survival has remained static over the past 20 years. This is due to the fact that little is known about the basic science mechanisms of the pathology of human osteosarcoma as well as the mechanisms of inherent drug resistance of OS. In particular lesser still is known regarding the role of tumor microenvironment that govern growth and progression of osteosarcoma in vivo. Recent clinical reports have demonstrated a negative correlation between tumor hypoxia and overall survival in osteosarcoma patients. In addition to the upregulation of HIFs, it has been shown that hypoxia can trigger an adaptive response such as the unfolded protein response (UPR) that allows tumor cells to avoid therapy-induced death.

Using in vitro experimental models of both the non-metastatic (SAOS-2) and metastatic (143-b) osteosarcoma cell lines, we now show that hypoxic signaling in these cells differentially activates a pro-survival mechanism via the up-regulation of the endoplasmic reticulum (ER) chaperone BiP and increased activation of the ER stress-activated eukaryotic translation initiator factor 2alpha kinase RNA-dependent protein kinase-like ER kinase (PERK) and ATF-6, markers of UPR allowing the cells to resist drug toxicity. Furthermore RNA interference and dominant-negative expression studies revealed that both ATF-6 and PERK signaling promote survival and drug resistance of OS cells,

Our findings shed light on the unknown mechanisms underlying chemotherapeutic drug resistance in osteosarcoma patients. We propose that hypoxic induction of UPR protects OS cells from stress insults, such as chemotherapy. Our research may lead to novel therapies that seek out and destroy the chemoresistant OS cells within the hypoxic core of tumors, thereby preventing survival and metastasis, and ultimately improving the chances of survival amongst OS patients.

Citation Format: Janine Faraj, Pooja Hingorani, Aparna R. Sertil. Differential activation of unfolded protein response in metastatic vs. non-metastatic osteosarcoma cells following hypoxia and chemotherapeutic stress. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3985. doi:10.1158/1538-7445.AM2014-3985
Abstract 3995: Quantitative protein profiling of tumor angiogenesis and metastasis biomarkers in mouse and human models
Authors: Lie Wen-Rong , Lipsey Jonathan , Warmke Tim , Yan Lin , Mistry Jehangir
Published: May 2015

Tumor and stromal cells secrete a variety of proteins acting as extracellular signals and creating a supportive microenvironment for tumor development, angiogenesis, and metastasis. We used the MILLIPLEX MAP cytokine/chemokine, cancer, angiogenesis, metastasis, MMP, bone metabolism, adipocyte, and cell signaling multi-pathway panels, based on the Luminex xMAP technology, to quantitatively evaluate the protein biomarker expression profiles on cultured mouse and human tumor cells and stromal cells as well as in vivo mouse and human tumor tissues. In the mouse system, biomarker profiling was performed to characterize secreted protein levels in Lewis Lung carcinoma (LLC) cells, B16.10 melanoma cells, b.End3 endothelial cells, and LLC tumors. The in vivo characterization, using primary tumors from mice bearing metastatic LLC, has identified differential expression of a panel of biomarkers, including amphiregulin (AREG), EGF, PECAM-1, KC, MCP-1, MIP-2, MMP-12, and resistin, between the high-fat (45% fat) and AIN-93G (15% fat) fed mice (p<0.05, 8 mice per group). These putative biomarkers may be involved in molecular mechanisms of the high-fat enhanced metastasis. Cell signaling pathway analysis revealed the activation of multiple pathways (JNK, p38, ERK, and STAT) in this mouse LLC model. In the human system, a heat map was generated to summarize the secreted protein signatures and the soluble receptor tyrosine kinases (RTK) profile on a series of the cultured human tumor cells (lung, brain, breast, prostate, liver, and colon) and 3 other human cell lines, HUVEC (endothelial), THP-1 (monocyte), and HFF-1 (fibroblast). Lastly, expression profiling of tumor and adjacent normal tissues from 2 patients with metastatic breast cancer was analyzed. Elevated levels of protein biomarkers, including MDC, IL-1RA, IL-8, IP-10, MCP-1, MIP-1α, OPG, OPN, SPARC, periostin, endoglin, PLGF, VEGF-A, MMP-1, MMP-7, MMP-9, and MMP-12, were detected in the breast tumors analyzed compared to their matched adjacent normal breast tissue. In summary, our results demonstrated that multiplexing using the Luminex xMAP bead-based platform for simultaneous quantitative expression profiling is a useful proteomic research tool, providing insights into biomarkers related to tumor progression and metastasis.

Citation Format: Wen-Rong Lie, Jonathan Lipsey, Tim Warmke, Lin Yan, Jehangir Mistry. Quantitative protein profiling of tumor angiogenesis and metastasis biomarkers in mouse and human models. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3995. doi:10.1158/1538-7445.AM2014-3995
Abstract 4: WISP-1 increases angiogenesis and VEGF-A expression in human oral squamous cell carcinomas
Authors: Tsao Ching-Wen , Tang Chih-Hsin , Chuang Jing-Yuan
Published: May 2015

Oral squamous cell carcinomas (OSCC) is most common metastasis to cervical lymph nodes by angiogenesis. Wnt-1 induced secreted protein 1 (WISP-1) is a cysteine-rich protein that belongs to the CNN family (CTGF / CYR61 / NOV). However, the effects of WISP-1 on angiogenesis and Vascular endothelial growth factor- A (VEGF-A) expression in human OSCC are not clear. First, we found that the expression of WISP-1 in human OSCC tissues were higher than in normal oral tissues. We also demonstrated that WISP-1 enhanced angiogenesis and VEGF-A up-regulation in human OSCC. WISP-1 induced VEGF-A expressio and angiogenesis through αvβ3 integrin, FAK, c-Src, EGFR, and ERK pathway. Knockdown WISP-1 decreased VEGF-A expression and also abolished human OSCC condition medium-mediated angiogenesis in vitro as well as angiogenesis effects in the chick chorioallantoic membrane (CAM) and matrix gel plug nude mice model in vivo. In addition, in the xenograft tumor angiogenesis model, knockdown of WISP-1 significantly reduced tumor growth and tumor-associated angiogenesis. We characterized an important role for WISP-1 which regulates angiogenesis by increasing VEGF-A expression in human OSCC via the αvβ3 integrin, FAK, c-Src, EGFR, ERK, and HIF1-α signaling pathway.

Citation Format: Ching-Wen Tsao, Chih-Hsin Tang, Jing-Yuan Chuang. WISP-1 increases angiogenesis and VEGF-A expression in human oral squamous cell carcinomas. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4. doi:10.1158/1538-7445.AM2014-4
Abstract 40: Association of Interferon-induced protein with tetratricopeptide repeats 1 and 3 expression with regional metastasis and drug resistance in oral squamous cell carcinoma
Authors: Yen Ai-Hsin , Chen Ying-Chen , Pidugu Vijaya Kumar , Liu Chung-Ji , Lee Te-Chang
Published: May 2015

Interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) gene family, consisting of 4 members (IFIT1, IFIT2, IFIT3, and IFIT5) in humans, is one of most responsive interferon-stimulated gene families. We have previously demonstrated that IFIT2 silencing activates atypical PKC signaling and induces epithelial-mescenchymal transition, cell migration, invasion, and metastasis of oral squamous cell carcinoma (OSCC) cells in cultured or animal models. These results implicate that IFIT2 is potentially a prognosis biomarker of patients with OSCC. In the present study, we further explore the roles of other IFIT1 family proteins involved in the regulatory mechanism of metastasis and drug resistance in OSCC cells. We determined the expression levels of IFIT1 gene family proteins in 82 patients with OSCC by immunohistochemical technique. Our present results revealed a close association between IFIT1 and IFIT3 protein expression in OSCC patient-derived tumor specimens (p < 0.001). However, the expression levels of IFIT1 or IFIT3 did not have a correlation with that of IFIT2 in tumor tissues. Intriguingly, IFIT5 was the only member of IFIT1 family displayed a well correlation with HPV E6 protein expression. Furthermore, we found that the expression levels of IFIT1 and IFIT3 were positively associated with age as well as regional lymph node metastasis status, whereas they were marginally and inversely associated with the differentiation features. We further demonstrated that IFIT1 and IFIT3 protein expression-induced by pre-treatment or co-treatment with IFN-β in Ca9-22 cells increased their resistance to Cisplatin and 5-flurouracil (5-FU). Silencing of IFIT1 and IFIT3 protein expression in SCC-25 cells by its specific shRNA, respectively, increased their susceptibility to Cisplatin and 5-FU. Since our present studies reveal that enhanced IFIT1 and IFIT3 expression associate with regional lymph node metastasis and drug resistance to Cisplatin and 5-FU, the roles of IFIT1 and IFIT3 expression associated with the clinicopathologic development in OSCC patients warrant our further investigation.

Citation Format: Ai-Hsin Yen, Ying-Chen Chen, Vijaya Kumar Pidugu, Chung-Ji Liu, Te-Chang Lee. Association of Interferon-induced protein with tetratricopeptide repeats 1 and 3 expression with regional metastasis and drug resistance in oral squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 40. doi:10.1158/1538-7445.AM2014-40
Abstract 4010: MicroRNA regulator of NMI: implications for breast cancer
Authors: Pruitt Hawley C. , III Jack W. Rostas , Metge Brandon J. , Devine Daniel J. , Bailey Sarah K. , Shevde Lalita A. , Samant Rajeev
Published: May 2015

Recent investigations by our laboratory have demonstrated that NMI expression is down-regulated in patient breast tumors, especially those displaying invasive/metastatic features. In breast cancer and melanoma cell lines, restored expression of NMI suppresses tumor growth. Furthermore, NMI negatively impacts TGFβ-driven epithelial-mesenchymal-transition (EMT) and alsometastatic invasion of breast cancer. Thus the mechanisms that regulate NMI expression are of potential interest to understand the etiology of breast tumor progression, invasion, metastasis and to identify potentially novel therapeutic targets.

MicroRNAs are critical regulators of gene expression. MicroRNA target prediction searches revealed that the NMI transcript could be targeted by the miR-29 family, a group of miRNAs with multiple roles in diverse cancer types. We hypothesized that miR-29 will negatively regulate NMI expression. Luciferase reporter assays using pMIR-REPORTTM confirmed the projected interaction of miR-29 in the seed region of the 3′ UTR of NMI. Manipulation of miR-29 levels in breast cancer cells by overexpression or silencing showed its inverse relationship with NMI expression. Invasive mesenchymal-like breast cancer cell lines showed elevated endogenous levels of miR-29 relative to tumorigenic/non-metastatic (epithelial like) breast cancer cell lines. Moreover, transient overexpression of miR-29 enhanced breast cancer cell invasion as measured by modified Boyden chamber assay and silencing miR-29 using anti-miRs decreased invasion. Investigations on the impact of miR-29 modulation on EMT revealed that increasing miR-29 levels in epithelial-like breast cancer cell lines caused 3D morphological changes indicative of invasive mesenchymal-like growth, while decreasing miR-29 levels from mesenchymal-like cell types using anti-miRs reversed the phenotype and the growth pattern resembled acinar structures. The significance of our observation was realized when we compared RNA isolated from patient derived breast cancer tumors and compared with RNA from matched adjacent normal breast specimens. Analysis using the McNemar's test showed a strong, inverse relationship between the expression of NMI and the miR-29. We contend that aberrant miR-29 expression may account for reduced NMI in some invasive breast cancers.

Citation Format: Hawley C. Pruitt, Jack W. Rostas III, Brandon J. Metge, Daniel J. Devine, Sarah K. Bailey, Lalita A. Shevde, Rajeev Samant. MicroRNA regulator of NMI: implications for breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4010. doi:10.1158/1538-7445.AM2014-4010
Abstract 4021: A kinome screen identifies SRPK1 to mediate breast cancer metastasis
Authors: Water Bob van de , Roosmalen Wies van , Devedec Sylvia L. , Meerman John , Foekens John , Martens John , Geiger Benny
Published: May 2015

Tumor cell migration plays key role in cancer cell dissemination and metastasis and is controlled by signaling-mediated cytoskeletal and cell-matrix adhesion remodeling. Using a phagokinetic track (PKT) assay with migratory H1299 cells, we performed a siRNA screen of almost 1,500 genes encoding kinases/phosphatases, adhesome-related and migration-related proteins, with the aim of identifying genes that determine tumor cell migration speed, and persistence. Thirty candidate genes, showing significant effect were validated in live tumor cell migration assays, and eight of those had a significant association with metastasis free survival in breast cancer patients, ITGB3BP, MAP3K8, NEK2, SHC1 and SRPK1 being most significant .

Further examination indicated that the expression of SRPK1, the most interesting gene identified in this screen , was highest in basal-like breast cancer and its expression level was correlated with poor disease outcome and preferred metastasis to the lungs and brain. In two independent breast tumor metastasis models, stable shRNA-based SRPK1 knockdown suppressed metastasis to distant organs, including lung, liver and spleen and inhibited focal adhesion reorganization. We provide here a comprehensive information resource on the molecular determinants of tumor cell migration and identify SRPK1 as a potential candidate drug target associated with breast cancer metastasis.

Citation Format: Bob van de Water, Wies van Roosmalen, Sylvia Le Devedec, John Meerman, John Foekens, John Martens, Benny Geiger. A kinome screen identifies SRPK1 to mediate breast cancer metastasis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4021. doi:10.1158/1538-7445.AM2014-4021
Abstract 4039: Exploration of a potent E-Selectin antagonist (GMI-1271) as a potential novel therapeutic for treating breast cancer metastasis to the bone and lung
Authors: Esposito Mark , Magnani John L. , Kang Yibin
Published: May 2015

The propensity of breast cancer to metastasize to the lung and bone with high frequency presents an attractive opportunity for specific therapeutic intervention. Targeting the initial steps of metastasis, such as survival in circulation, arrest, and extravasation, may constitute a more effective approach than treatment during the later steps. E-Selectin has been implicated as a pro-metastatic endothelial receptor which mediates the initial stages of tumor cell arrest and extravasation to the lung, liver and bone. We examined the ability of the potent E-selectin antagonist GMI-1271 (Kd = 0.54 μM) to inhibit E-selectin binding to lung and bone metastatic cells. Flow cytometry and ELISA analysis was used to evaluate the effects of GMI-1271 in vitro while treatment of bone and lung metastasis with GMI-1271 during the initial stages of metastatic seeding was used in xenograft models. Results demonstrated that metastatic breast cancer cells express carbohydrates ligands recognized by E-selectin as determined by interaction with E-selectin-Fc chimeric protein and the antibody HECA-452. GMI-1271 could significantly reduce metastatic breast cancer cell binding to E-selectin in vitro. Treatment of mice following intracardiac injection resulted in a significant reduction of mortality (HR = 0.29, p = 0.044) while treatment following tail vein injection demonstrated a directional increase in survival (p = 0.061 and p = 0.055). These results designate GMI-1271 as a potential therapeutic for treatment of late stage breast cancer.

Citation Format: Mark Esposito, John L. Magnani, Yibin Kang. Exploration of a potent E-Selectin antagonist (GMI-1271) as a potential novel therapeutic for treating breast cancer metastasis to the bone and lung. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4039. doi:10.1158/1538-7445.AM2014-4039
Abstract 4051: Prion protein and its ligand STI1/HOP modulate migration and invasion of cell lines derived from colorectal tumors
Authors: Lacerda Tonielli S. , Dias Marcos Vinicios S. , Baptistella Antuani Rafael , Giudice Fernanda S. , Roz Bruna R. , Rodrigues Iara S. , Martins Vilma Regina
Published: May 2015

The colorectal cancer is the fourth most common type of cancer in the world. The search for potential molecular targets for therapy has led to the Cellular Prion Protein (PrPC) as a possible candidate. Its activity depends on the interaction with the secreted form of the stress inducible protein 1 or heat shock organizing protein (STI1/HOP). Previous results indicated the importance of the PrPC-STI1/HOP complex in proliferation of glioblastomas and invasion of melanomas. Therefore, this work aims to understand how PrPC-STI1/HOP complex can influence the migration and invasion of WiDr and HCT8 colorectal tumor cells. PrPC is equally expressed at the cell surface of both cell lines. Cell migration assays were performed using boyden chambers and recombinant STI1/HOP as chemical attractant. For invasion experiments cells were seeded on boyden chambers covered with matrigel and treated with STI1/HOP. The results showed that when stimulated with recombinant STI1/HOP, both tumor cell lines showed higher migration and invasion potentials than non-treated cells or those treated with a STI1/HOP deletion mutant for the PrPC binding domain (STI1Δ230-245). Moreover, migration and invasion mediated by PrPC-STI1/HOP complex were blocked by a STI-1/HOP (230-245) peptide, which mimics STI1 binding site to PrPC or also by a neutralizing anti-PrPC antibody. STI1/HOP is secreted in the conditioned medium (CM) within extracellular microvesicles (MVs) and WiDr cells secreted more MVs and STI1/HOP than HCT8 cells. Remarkable, HCT8 cells treated with CM or isolated MVs from WiDr cells show a higher migration than that induced when the former cells were treated with their own CM or when WiDr cells were treated with CM or MVs derived from HCT8 cells. Together, these results indicate that the PrPC-STI1/HOP complex mediate migration and invasion of colorectal tumor cells and suggest that cellular migration is dependent on the concentration of secreted MVs. The role of STI1/HOP present in MVs on cell migration is still under investigation.

Citation Format: Tonielli S. Lacerda, Marcos Vinicios S. Dias, Antuani Rafael Baptistella, Fernanda S. Giudice, Bruna R. Roz, Iara S. Rodrigues, Vilma Regina Martins. Prion protein and its ligand STI1/HOP modulate migration and invasion of cell lines derived from colorectal tumors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4051. doi:10.1158/1538-7445.AM2014-4051
Abstract 4052: Targeting PDK1 in breast cancer: kinase-dependent regulation of tumor growth and kinase-independent regulation of cell migration and invasion
Authors: Gagliardi Paolo Armando , Blasio Laura di , Seano Giorgio , Sessa Roberto , Puliafito Alberto , Bussolino Federico , Primo Luca
Published: May 2015

3-Phosphoinositide Dependent Kinase 1 (PDK1) is the kinase that phosphorylates the activation loop of several protein kinases such as Akt, PKC or SGK. In breast carcinomas PDK1 was found frequently overexpressed and this correlates with a more aggressive phenotype.

This study is aimed to understand the role of PDK1 in breast tumor progression and the molecular mechanisms involved.

Using stable gene silencing and kinase inhibitors, we studied PDK1 role in different in vitro and in vivo assays including anchorage independent growth, anoikis, xenografts, migration and invasion assays.

In our study we found that PDK1 promotes resistance to anoikis, anchorage independent growth and tumor growth in breast cancer cell lines through the phosphorylation of its substrates, since the kinase-dead mutant abrogates all the PDK1-mediated effects. For this reasons either PDK1 gene silencing or its kinase inhibition with small compounds are sufficient to promote anoikis, to reduce anchorage independent growth and tumor growth.

We also found that PDK1 silencing is able to reduce, while its overexpression to increase, cell migration and invasion in mammary cells. Surprisingly, the regulation of cell migration is operated in a kinase independent manner. In fact PDK1 kinase inhibitor failed to recapitulate the effects of gene silencing in cell migration. The molecular mechanism, underlying this unconventional kinase-independent regulation of cell migration, involves the binding of PDK1 to Myotonic Dystrophy kinase-related Cdc42-binding Kinase α (MRCKα). This binding increases MRCKα kinase activity on Myosin Light Chain-2 (MLC2) and on myosin phosphatase target subunit 1 (MyPT1). Both these phosphorylations increase of non-muscular myosin activity, which is required to pull forward the cell body during directional migration and invasion.

In summary, our findings unveil that PDK1 regulates tumor growth, survival in absence of adhesion, resistance to anoikis, but also migration and invasion of breast cancer cells by two different mechanisms. These findings should be taken in account for a rational targeting of PDK1 in cancer.

Citation Format: Paolo Armando Gagliardi, Laura di Blasio, Giorgio Seano, Roberto Sessa, Alberto Puliafito, Federico Bussolino, Luca Primo. Targeting PDK1 in breast cancer: kinase-dependent regulation of tumor growth and kinase-independent regulation of cell migration and invasion. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4052. doi:10.1158/1538-7445.AM2014-4052
Abstract 4058: Acid-sensing ion channel 1a (ASIC1a) contributes to cell proliferation and migration caused by acidic environment in human glioma cells
Authors: Liu Mingli , Guo Alyssa (Aihui) , Ghiathi Christopher , Xiong Zhigang
Published: May 2015

Due to an increased metabolic rate, an acidic environment is commonly observed in tumor tissues. The acidic environment likely has a significant impact on the viability and proliferation of tumor cells. Acid-sensing ion channel (ASIC)1a is the key proton receptor expressed in nervous systems, which regulates acidosis-induced neuronal injury in various neurological diseases, such as ischemic stroke. Glioblastomamultiforme (GBM) is the most common and aggressive primary brain tumor in adults with median survival time of 14.6 months. Since GBM are characterized by chronic hypoxia and poor perfusion that is likely to result in acidosis, acidosis may influence the growth behavior and response to cytotoxic therapy. In the present study, we examined the effects of acidosis on growth and migration of cultured human malignant glioma cells A172, and the role of ASIC1a in the cell death induced by extracellular acid. Methods: 1) We detected ASIC1a mRNA using reverse-transcription polymerase chain reaction (RT-PCR). 2) We then determined the presence of ASIC1a in A172 cells using electrophysiological recording (patch-clamp technique). 3) We assessed the effects of acidic environment on cell proliferation and maintenance of clonogenic capability using MTT assay and colony formation assay respectively. 4) Finally, we examined the effects of acidic environment on glioma cell migration by transwell invasion assay. Results: 1) RT-PCR detected the presence of ASIC1a and ASIC4 mRNA in A172 cells. 2) The inward current was recorded when treated with pH5.0 solution indicating the expression of functional ASIC1a in A172 cells. 3) Acidic environment decreased the number of surviving A172 cells, the colony formation, and also inhibited A172 cell migration. 4) Inhibition of ASIC currents by nonspecific inhibitor of ASICs amiloride or specific inhibitor PcTX1 protected A172 cell from death. Conclusions: Extracellular acid reduces proliferation and migration in glioma cells. Extracellular acid induces death of glioma cells via ASIC1a mechanisms. These results indicate the presence of ASIC1a in glioma cells and may play significant role in glioma progression. This work was supported by the National Institutes of Health grant numbers NIH R01NS047506, R01NS066027, UL1 RR025008, U54 RR026137, AHA 0840132N, and ALZ IIRG-10-173350.

Citation Format: Mingli Liu, Alyssa (Aihui) Guo, Christopher Ghiathi, Zhigang Xiong. Acid-sensing ion channel 1a (ASIC1a) contributes to cell proliferation and migration caused by acidic environment in human glioma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4058. doi:10.1158/1538-7445.AM2014-4058
Abstract 4060: Androgen receptor is involved in the migration and invasion of triple-negative breast cancer cells
Authors: ZHENG Qiao-dan , Wu Wenyuan , Huang Cai
Published: May 2015

Triple-negative breast cancer(TNBC)is a major world health problem, accounting for more than 15% of breast cancer new cases each year. The 5-year survival rate for TNBC is approximately 54.5% and death usually occurs as a result of local invasion and regional metastases. A complete understanding of the molecular mechanisms that control the aggressive behavior of triple-negative breast cancer cells is essential in order to develop targeted therapies to manage the disease. Recent evidence has suggested a role of Androgen receptor (AR)in cellular motility and invasion. In this present study we examined the role of AR on the migration and invasion of triple-negative breast cancer cells. Detect the AR levels in 7 cell line of breast cancer cells including MCF-7, MDA-MB-157, MDA-MB-231, MDA-MB-435, MDA-MB-468, MDA-MB-549, HS-578T by Q-PCR, we demonstrate that AR is highly express in triple-negative breast cancer cells; shRNAs specific to AR have been generated and delivered into human breast cancer cells by lentiviral vectors. We showed that the total levels of AR can be decreased by AR shRNA and inhibit MDA-MB-231 cellular motility and large focal adhesion (FA) formation, the cell migration was determined by time-lapse cell migration assays, depletion of endogenous AR significantly reduced the velocity and directionality of cell migration, and strongly inhibited the net distance of cell migration, indicating that AR modulates cell migration by controlling the velocity and directionality of migration. We also showed that AR activation and the migration and invasion of MDA-MB-231 , MDA-MB-435, MDA-MB-468 can be strongly inhibited with commercially available AR blocker Bicalutamide by cell migration and invasion assay. These results suggest that AR in breast cancer cells is required for cellular migration and invasion.

Citation Format: Qiao-dan ZHENG, Wenyuan Wu, Cai Huang. Androgen receptor is involved in the migration and invasion of triple-negative breast cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4060. doi:10.1158/1538-7445.AM2014-4060