bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4662: Genetic variations in the cell-cycle pathway and survival of patients with advanced non-small cell lung cancer receiving platinum-based chemotherapy
Authors: Yin Jikai , Wu Xifeng , Lu Charles , Chang Joe , Hildebrandt Michelle , Stewart David , Spitz Margaret R. , Gu Jian
Published: February 2011

Background: Platinum-based chemotherapy is the main choice of treatment for patients with advanced Non-small cell lung cancer (NSCLC). Platinum agents target DNA preferentially for cytotoxicity, which will lead to DNA damage and cell-cycle arrest. We hypothesize that genetic variations in cell cycle pathway may modulate response to platinum-based chemotherapy. In this study, we utilized a comprehensive pathway-based approach to identify genetic variations in the cell-cycle pathway as predictors of overall survival in NSCLC patients treated with platinum-based chemotherapy. Material and Methods: We genotyped 633 polymorphisms from 105 cell cycle-related genes in 598 advanced NSCLC cases who received first-line platinum-based chemotherapy. All genotypes were analyzed for association with overall survival and evaluated for potential cumulative and gene-gene interactions that further modulated overall survival. Multiple statistic analyses were applied, including individual SNP analysis, cumulative genotype analysis, haplotype analysis, survival tree analysis and Kaplan-Meier survival analysis. Results: 65 SNPs from 40 cell-cycle genes were identified that were significantly associated with NSCLC overall survival (p<0.05) individually. We found that increasing numbers of potential high-risk alleles in any cell-cycle checkpoint would lead to increased risk of death. Compared with other checkpoints, G1/S checkpoint showed more significant associations with patients’ survival. For the combined analysis of unfavorable genotypes in G1/S checkpoint, compared with the referent group (≤20 unfavorable genotypes), the HRs for patients carrying 21-23 and 24-32 unfavorable genotypes were 1.94(95%CI 1.47-2.56) and 3.00(95%CI 2.33-3.85), respectively (p for trend < 0.0001), and also a significant reduction of median survival time from 24.05 to 9.18 months (log-rank p < 0.0001). Heplotype analysis revealed that a TP73 haplotype (GTGTTA) was significantly associated with increased risk of death (p=0.00026). Survival tree analysis of overall cell-cycle pathway indicated G1/S checkpoint genes categorized patients into subgroups with different survival pattern. Conclusions: These results suggest that G1 and G1/S checkpoint genetic variations affect survival of advanced NSCLC patients treated with platinum-based chemotherapy and that there are gene-dosage effect and gene-gene interactions in modulating clinical outcomes.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4662.
Abstract 4664: Fast analysis and quantification of collagen fibril alignment in the tumor microenvironment in an MNU-induced model of mammary carcinogenesis
Authors: Ericson Marna E. , Gallus Nathan , Grubbs Clinton J. , Bode Ann , Lubet Ronald
Published: February 2011

We have developed a one-day imaging acquisition and analysis protocol to quantify orientation of the collagen fibrils at the boundary of a mammary tumor lobule in a tissue biopsy. We reported that the degree of radially-aligned collagen fibrils in the tumor-stromal interface of rodent mammary tumor tissue is an indicator of chemotherapeutic potential of the EGFr inhibitor, gefitinib (Ericson et al. 2009 In: Proc Annu Meet AACR; Abstract nr 5002).

The MNU-induced model of mammary carcinogenesis was used to develop image capture and analysis protocols. Small, 1-mm punch biopsies, were acquired from female rats with palpable (app. 250 mm2) tumor masses. Biopsy tissues were fixed and cut into 150-micron thick sections. No staining was necessary as fibrillar collagen, a hyperpolarizable structural protein, is readily detected using second harmonic generation imaging (SHG). Using this technique, the collagen fibrils can be imaged using fresh ex vivo, fixed, or paraffin-embedded tissue.

Multiple SHG images of adjacent fields of view (FOV) were acquired at 200X magnification over the entire 1-mm biopsy section. Each FOV image is a stack of 14 five-micron optical sections, acquired in the Z-plane of the sample. This is an automated process using a programmable stage allowing for fast 3D data acquisition over the entire section. We developed Photoshop and Image J plug-ins to automate the projection of the individual optical Z-stacks and subsequent image stitching of all the projections. Tumor/stromal boundaries were then identified by the operator. The angle of divergence to the tangent at the tumor interface for all fibrils was quantified. Collagen fibrils running parallel to the tangent are at zero degrees while fibrils perpendicular to the tangent are at 90 degrees. This analysis can be completed in a single day.

More perpendicular collagen orientation at the tumor/stromal interface may be indicative of a fast-growing aggressive tumor or enhanced ability of the tumor to invade surrounding tissue whereas predominant parallel collagen orientation could reflect increased encapsulation of the tumor mass which would then discourage further tumor growth or as our work has shown, the collagen orientation may serve as an indicator of responsiveness to chemotherapy. The development of an indicator/test for chemotherapeutic efficacy would be a useful tool for drug design or in creation of an individualized patient treatment plan.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4664.
Abstract 4668: Pro-inflammatory lipid PAF may induce malignant potential via PAFR and phospho-FAK/STAT pathway in BRCA1-positive tubo-ovarian epithelial cells
Authors: Zhang LiFang , Wang Dan , Edwards Dale , Aponte Margarita , Qiu Weiliang , Jin LianJin , Seethapan Vanitha , Vitonis Allison , Wang Jian Liu , Mok Samuel , Crum Christopher , Cramer Daniel W. , Ye Bin
Published: February 2011

The loss of BRCA1 function may induce profound transcriptional changes, phenotypic reprogramming and ultimately, generate genetically unstable tumor-initiating cells, which significantly increase the risk of breast and ovarian cancer. The molecular targets and pathways involved in early events of malignant transformation of at-risk epithelium has not been well investigated, particularly the effects of chronic inflammation. In an earlier study, we demonstrated that platelet activating factor (PAF), a potent pro-inflammatory mediator, plays a significant role in ovarian cancer progression and invasion (Cancer Research, 68:5839, 2008). Here, we extend our finding to show that PAF receptor is over expressed in BRCA1-mutated (BRCA1+) human ovarian surface epithelial cells (HOSE) by western blot and in distal fallopian tube (fimbria) epithelial cells by immunohistochemistry. In addition, PAF induced cell apoptosis only in wild type HOSE cells, and anti-apoptosis in BRCA1+ ovarian surface epithelial cells. A kinase microarray revealed that phosphor-STAT1/3/4/6 was simultaneously activated by 100 nM PAF treatment in BRCA1+ ovarian cancer cells. Induction of phosphor-STAT1 associated with PAFR and BRCA1-mutation was further confirmed in three BRCA1+ HOSE cell lines, but not in normal (HOSEE6E7) or malignant (OVCA429) ovarian epithelial cells. Co-immune precipitation revealed that PAF/PAFR coordinately activates FAK through dephosphorylation and STAT1 through phosphorylation only in BRCA1+ ovarian epithelial cells. These findings strongly suggest that potent inflammatory mediators (i.e. PAF) may play a significant role to induce malignant transformation in BRCA1-mutant ovarian epithelial cells through the FAK-STAT pathway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4668.
Abstract 467: Effects of Monascus-fermented rice extract on malignant cell associated neovascularization and intravasation by chicken embryo chorioallantoic membrane model
Authors: Ho Bing-Ying , Wu Yao-Ming , Hsu Ya-Wen , Hsu Li-Chuan , Kuo Yao-Haur , Chang King-Jen , Pan Tzu-Ming
Published: February 2011

The traditional Chinese medicine, “Hong-Qu”, also call “red mold rice” in the United States and Europe, has been identified and used for the treatment of blood stasis, a disorder related to hyperlipidemia and atherosclerosis. In addition to its value in improving metabolism syndrome, extracts from Monascus fermented rice inhibited proliferation of various cancer cells in vitro and in vivo have demonstrated previously. The objective of the current study was to examine correlation of red mold rice ethanol extract (RMRE) on the process of angiogenesis, invasion and metastasis during tumor progression. In the current study, we examined the inhibitory effects of SW480 and SW620 human colorectal carcinoma cells by RMRE using in vitro MTT assay. RMRE significantly inhibited the proliferation of SW480 and SW620 cells in dose- and time-dependent manner. However, human umbilical vein endothelial cells (HUVECs) showed a minor reduction effect of cell viability up to 20 µg/mL of RMRE for 48 hours using crystal violet staining assay. The capillary-like network morphology was observed following 20 ng/mL VEGF or SW620 culture-conditional medium (CM) addition. Nevertheless, capillary-like tube formation have failed to show when RMRE participation. Moreover, SW620 cells could growth in Matrigel grafts and spontaneously intravasation from the upper to the lower of chick embryo chorioallantoic membrane (CAM) model, which detected human Alu genomic DNA by PCR amplification from the lower CAMs at RMRE-untreated group. The percentage of neovascularization was increased 75.3±11.6% by SW620 cells onplant with Matrigel grafts on the CAM model. However, addition of RMRE significantly reduced CAM neovascularization in dose-dependent effect. Finally, we have finding that RMRE effectively decreased activity of matrix metalloproteinase (MMP)-7 determined by RT-PCR, Western blotting, and casein zymography assays. In summary Monascus fermented products exerted a potent effect on tumor growth and activation, suggesting that it may useful serves as a supplementary agent in adjuvant cancer therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 467.
Abstract 4671: Tumor Suppressive Role of HOP (homeobox only protein) Gene in Gastric Cancer
Authors: Yamashita Keishi , Ooki Akira , Katoh Hiroshi , Sidransky David , Watanabe Masahiko
Published: February 2011

Background: Homeobox only protein (HOP) is an unusual homeobox gene encoding the 3 spliced transcript variants, among which the only HOP-β promoter harbors CpG islands and inactivated by methylation in esophageal and gastric cancer (Yamashita K et al., Cancer Cell, 2002, Kim MS and Yamashita K et al., Cancer Res., 2006, Yamashita K et al., Mol. Cancer Res., 2009). HOP acts as a novel tumor suppressor gene, but remains elusive for functional role in human cancer.

Methods: Using bisulfite sequencing analysis and quantitative-methylation specific PCR (Q-MSP), HOP-β methylation status was evaluated in 80 pairs of gastric cancer and the corresponding normal tissues, further validated in 90 advanced gastric cancer. Transfection of HOP into gastric cancer cell line GCIY was done to examine whether it actually has a tumor suppressive function.

Results: By Q-MSP, HOP-β hypermethylation was found in 84% (67/80) primary tumor tissues and 10% (8/80) corresponding normal tissues. When the cut-off value was set at 11.28, the HOP-β methylation was a potent prognostic factor (P=0.0006), which was further validated in the independent 90 advanced gastric cancer (P=0.009). Multivariate analysis revealed that HOP-β methylation could be independent prognostic factors. HOP-β expression was silenced by the promoter methylation in all gastric cancer cell lines tested; expression could be restored by treatment with methyltransferase inhibitor. Moreover, reintroduction of HOP-β gene repressed cell growth and invasion.

Conclusion: HOP-β promoter methylation is a frequent and cancer-specific event in gastric cancer. Quantitative assessment of HOP-β methylation has great clinical potential as a marker of tumor aggressiveness and may thus facilitate treatment planning in patients with advanced gastric cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4671.
Abstract 4673: Myopodin methylation is a prognostic marker in patients with T1G3 bladder cancer
Authors: Alvarez Miguel , Cebrian Virginia , Fernandez Jesus , Fresno Florentino , Escaf Safwan , Sanchez-Carbayo Marta
Published: February 2011

BACKGROUND: Urothelial carcinomas presenting T1G3 non muscle-invasive lesions are high-risk tumors because of the high recurrence and progression to muscle invasive disease rates. The Bacillus of Calmette-Guérin (BCG) is a standard treatment to reduce tumor recurrence and delay progression of high-risk non-muscle invasive bladder tumors. However, it is not clear yet which T1G3 patients are more prone to display a more aggressive clinical behavior or be susceptible to respond to BCG.

OBJECTIVE: The aim of this study was to evaluate the role of myopodin methylation as a clinical outcome prognosticator and predictive biomarker for BCG response in patients with T1G3 bladder tumors.

DESIGN, SETTING, AND PARTICIPANTS: The methylation status of myopodin was analyzed on tumor specimens belonging to 170 patients with T1G3 non-muscle invasive bladder cancer, being a subset of them undergoing BCG treatment (n=108).

MEASUREMENTS: Myopodin methylation was assessed by methylation-specific polymerase chain reactions. Recurrence, progression into muscle invasive tumors and disease-specific overall survival rates were analyzed using competing risks regression analysis.

RESULTS: Among the 170 cases analyzed, 72 of them recurred (42.4%), 36 progressed (21.2%), and 24 died of the disease (14.1%). Univariate and multivariate survival analyses revealed that myopodin methylation was significantly associated with an increased recurrence rate (p=0.004), progression (p=0.002), and shorter disease-specific overall survival (p=0.020). Interestingly, in a subset of cases treated with BCG, myopodin methylation was also related to an increased recurrence rate (p=0.011), progression (p=0.030), and a shorter disease-specific overall survival (p=0.028).

CONCLUSIONS: Epigenetic analyses revealed that myopodin methylation was associated with the clinical outcome of patients with T1G3 tumors. Myopodin methylation was related to tumor aggressiveness in patients with T1G3 disease undergoing BCG treatment. Myopodin methylation distinguished patients responding to BCG from those who may require a more aggressive therapeutic approach.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4673.
Abstract 4674: Receptor for advanced glycation endproducts (RAGE) regulates ß-catenin function and chemosensitivity in pancreatic cancer cells
Authors: Kang Rui , Tang Daolin , Lotze Michael T. , Zeh Herbert J.
Published: February 2011

β-catenin is a key downstream effector in the Wnt signaling pathway. It is implicated in two major biological processes in vertebrates: early embryonic development and tumorigenesis. The receptor for advanced glycation endproducts (RAGE), a member of the immunoglobulin superfamily of cell membrane receptors, is encoded within the MHC Class III Region. It interacts with various ligands including high mobility group protein 1 [HMGB1], certain S100 protein family members, and advanced glycation endproducts and is involved in cancer, inflammation, diabetes and Alzheimer disease. To evaluate whether treatment with RAGE short hairpin RNA (shRNA) increases chemosensitivity in pancreatic cancers involves β-catenin signaling, we first examined the expression level of β-catenin and its target genes. In normal states, β-catenin and its target genes cyclinD1, c-myc, and Nr-CAM but not L1CAM were decreased when RAGE was depleted in the murine Panc02 cell line by western blot and immunofluorescence analysis. Importantly, RAGE deletion enhanced the suppressive effects of chemotherapy on the expression of β-catenin and its target genes. To evaluate the effects of β-catenin on chemosensitivity in pancreatic cancer cells, we used cells in which the expression of RAGE and/or β-catenin was suppressed by shRNA. Knockdown of β-catenin increased chemosensitivity in pancreatic cancer cells and these effects were enhanced when both β-catenin and RAGE were knocked down. Notably, knockdown of β-catenin had no effect on the commitment of cells to enhanced autophagic flux when assessed by evaluation of microtubule-associated protein 1 light chain 3 (LC3) punctae formation. However, knockdown of β-catenin did significantly increase apoptosis when assayed by increased activity of caspase 3 and cleavage of caspase3. Taken together, these results suggest that RAGE regulates β-catenin transcription function and chemosensitivity in pancreatic cancer cell lines.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4674.
Abstract 4675: Lack of evidence for XMRV infection in German prostate cancer patients
Authors: Krause Hans , Hohn Oliver , Miller Kurt , Kurth Reinhard , Bannert Norbert
Published: February 2011

A novel gammaretrovirus named xenotropic MuLV-related retrovirus (XMRV) has been recently identified with 40 % prevalence in prostate tumor samples from US American patients carrying a homozygous R462Q mutation in the RNASEL gene. This mutation impairs the function of the innate antiviral type I interferon pathway and is a known susceptibility factor for prostate cancer. In subsequent studies, analyzing small European cohorts of prostate cancer patients a less significant prevalence and correlation of the QQ-phenotype with XMRV has been found or even a complete absence of viral traces was reported. A second independent report from the US found XMRV DNA in 6% and XMRV protein expression in 23% of prostate cancers. Contrary to the initial report the XMRV infection was not associated with this common variant of RNASEL and predominantly observed in malignant epithelial cells.

We aimed to determine the prevalence of XMRV in prostate cancer patients who had undergone radical prostatectomy at the Charité hospital in Berlin, Germany between 2000 and 2006. To validate the association of the XMRV infection with the R462Q polymorphism, we first genotyped our 589 prostate tumour samples for the G-to-A transition at nt1385 of the RNASEL gene. Using nested PCR and RT-PCR approaches, DNA and RNA samples of prostate cancer (PCa) patients with varying degrees of malignancy were screened for the presence of XMRV specific gag sequences. Furthermore 146 sera samples from PCa patients were tested for XMRV Gag and Env antibodies with a newly developed ELISA assay.

Genotyping our patients revealed the presence of the minor allele (QQ genotype) in 76 samples (12.9 %). Although using highly sensitive PCR-based detection methods we could not detect XMRV specific sequences either at the DNA or RNA level. Consistent with this result, all analysed sera from prostate cancer patients did not contain XMRV specific antibodies.

Our results indicate a complete absence or at least a much lower prevalence of XMRV in prostate tumour patients in Germany.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4675.
Abstract 4677: Irinotecan plus anthracycline - based combination chemotherapy in advanced solid tumors
Authors: Rebollo Joseba , Sureda Manuel , Gonzalez-Manzano Ramon , Balenzuela Belen , Brugarolas Antonio
Published: February 2011

Irinotecan and Anthracyclines (Topoisomerase I and II inhibitors, respectively) are widely used as antitumoral agents in cancer treatment. However, no data have been reported when used in combination.

In this study we report the activity of biweekly Irinotecan (150 mg/m2) plus Anthracycline (Doxorrubicin 30 mg/m2 or Pegylated Doxorrubicin 20 mg/m2) - based combination chemotherapy in some previously treated advanced solid tumours.

From June ’04 to June ’09, eighteen patients (pt) have been treated. Median age has been 64 y (range 28-84 y). Eight pt had ovarian cancer, 8 pt non-small-cell lung cancer (NSCLC) and 2 pt gastric cancer. Therapy has been administered as a second line treatment in 11 pt and as a third and further treatment in 7 pt. Four pt have received radiotherapy previously.

Toxicities have been leucopenia (3 pt grade III-IV, 6 pt grade I-II), mucositis (6 pt grade III-IV, 5 pt grade I-II), diarrhea (2 pt grade III) and hand-foot syndrome (2 pt grade II). One pt died of neutropenic sepsis after the first dose of chemotherapy.

Seven out of 16 (44%) pt presented objective response: 4/7 ovarian cancer pt, 2/7 NSCLC pt and 1/2 gastric cancer pt. Two pt are not evaluable for response (one pt without evaluable disease and 1 pt early toxic death). Median Progression Free Survival has been 4 mo (range 1-13 mo).

Irinotecan plus Anthracycline - based combination chemotherapy is an active treatment in some solid tumours and it is well tolerated. Most of the toxic events were related to heavily pre-treated pt. It is infrequent to observe objective good quality responses in first line therapy resistant tumours. For this reason, we purpose there is a synergistic effect based on the known mechanisms of both drugs (Topoisomerase I and II inhibition) and an early-use first line therapy is recommended with this combination. This combination should be tested in larger trials to confirm antineoplastic activity.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4677.
Abstract 4680: Correlation between Wnt pathway and colorectal cancer recurrence using protein, mRNA and microRNA analysis
Authors: Coudry Renata A. , Martins Mila O. , Coutinho Cláudia M C , Buim Marcilei , Carvalo Kátia C. , Santos Erika M. , Brentani Helena
Published: February 2011

BACKGROUND: Wnt signaling pathway has a fundamental role in colorectal carcinogenesis. However, few studies have focused in the association of this pathway and the risk of postoperative recurrence. Besides microRNAs repress protein translation through binding to targets mRNAs and have shown association with specific targets. The main goal of this study was to evaluate several members of Wnt signaling pathway (Wnt protein family, proteins from β-catenin/APC complex, transcription factors and target proteins) and CRC recurrence. We also would like to assess association of microRNAs with specific Wnt targets or the direct implication of those small non coding RNAs with CRC recurrence. METHODS: The expression of Wnt proteins and mRNAs were assessed using immunohistochemistry and Real Time-PCR techniques, respectively. We also studied 381 microRNAs using Low Density TaqMan Array. This study was performed in samples from patients presenting stages I and II CRC and comparing those who had recurrence to those who did not. All patients undergone surgery with curative intention from 2000 to 2006 in A. C. Camargo Hospital. Statistical analyses used in this study were Chi-square, exact Fisher's test and T Paired. RESULTS: Regarding the protein expression and gene expression of Wnt signaling members only c-Myc showed statistically significant association with recurrence (p=0,001 and p=0,048, respectively). Regarding the analysis of 381 microRNAs, a statistically significant association was observed for miR-197 (p=0,03), where the group of patients who did not had recurrence demonstrated higher expression levels of this microRNA when compared to the group of patients who developed tumor recurrence. CONLUSION: Increased levels of c-Myc expression and low levels of miR-197 in the tumor may be used at the time of primary removal for defining patients at risk for tumor recurrence. miR-197 may regulate indirectly the expression of c-Myc. However, further experiments will be required to better evaluate this hypothesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4680.
Abstract 4685: Multiplexed methylation profiles of urinary DNA for non-invasive bladder cancer diagnostics
Authors: Cabello Maria Jose , Grau Laura , Orenes-Piñero Esteban , Franco Noreli , Alvarez Miguel , Blanca-Pedregosa Ana , Heredero Oscar , Fernandez Jesus , Palacios Alberto , Lopez-Beltran Antonio , Sanchez-Carbayo Marta
Published: February 2011

Changes in DNA methylation of crucial cancer genes including tumor suppressors can occur early in carcinogenesis, being potentially important early indicators of cancer. The objective of this study was to examine a multiplexed approach to assess the methylation of tumor suppressor genes as urinary diagnostic markers for bladder cancer. A multicandidate probe panel interrogated DNA for aberrant methylation status in 25 tumor suppressor genes using a methylation-specific multiplex ligation-dependent probe amplification assay (MS-MLPA). Initial analyses in bladder cancer cell lines (n=14), and fresh-frozen primary bladder tumor specimens (n=31) supported the panel of genes selected to be altered in bladder cancer. MS-MLPA was then optimized for its application in body fluids using two sets of urinary specimens (n=146) including bladder cancer patients (n=96) and controls (n=50). Genes previously reported to be methylated such as BRCA1, CDH13 and RARB, were confirmed to be methylated in the three types of samples analyzed. WT1 and PAX5A were identified as novel tumor suppressor candidates methylated in bladder cancer specimens. The most frequently methylated genes in the bladder tumors analyzed were BRCA1 (71.0%), WT1 (38.7%), and RARB (38.7%), being WT1 methylation significantly associated with tumor stage (p=0.011). BRCA1, WT1 and RARB were also as the most frequently methylated genes in two independent training and validation sets of urinary specimens (n=146). ROC curves analyses revealed significant diagnostic accuracies over 70% in both sets of urinary specimens for BRCA1, RARB, WT1, PAX5A and CDH13. In summary, the novelty of this report deals with applying a multiplexed methylation technique for bladder cancer, and also in body fluids. MS-MLPA represents a promising methylation profiling tool for histopathologic stratification of bladder tumors, and diagnosis using urinary samples. This latter is clinically relevant by offering a non-invasive strategy for the clinical management of patients affected with uroepithelial neoplasias

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4685.
Abstract 4686: MAL is down-regulated in oral squamous cell carcinoma and suppresses proliferation, invasion, tumorigenicity of oral cancer cells, in vitro and in vivo
Authors: Chen Wantao , Cao Wei , Zhang Ping
Published: February 2011

Purpose: The molecular mechanisms involved in oral squamous cell carcinoma (OSCC) still remain unclear to date. Our previous study revealed that MAL was down-regulated in the tissues of oral squamous cell carcinomas tested by cDNA microarray analysis. In order to identify the function of MAL, the effects on the proliferation, invasion and apoptotic potential in oral cancer cell lines were investigated, in vitro and in vivo.

Methods: Expression levels of MAL mRNA in oral squamous cell carcinoma (OSCC) tissues and oral cancer cell lines were examined by semi-quantitive reverse transcription (RT)-PCR and quantitive Real-time PCR. OSCC cell line, Tca cells, were transiently and stably transfected with the full-length MAL cDNA sequence or the empty expression vector pcDNA3.1. Transient and stable gene transfers were verified by Western blot analysis and RT-PCR. Subsequently, WST-8 assay, colony-formation assay, and flow cytometry were used to assess the effects of MAL on the proliferation of Tca cells. Furthermore, PI staining, 7-AAD and annexin V staining and detecting the level of cleavage PARP protein were used to examine apoptosis of Tca cells by MAL gene transfer. Finally, Invasion and tumorigenicity potential of MAL-expressing or MAL-nonexpressing Tca cells was investigated by invasion assay in vitro and tumorigenicity assay in vivo. All statistical tests were two-sided.

Results: Expression of MAL was decreased in 86.7% (26/30) OSCC tissues compared with the matched normal tissues by semi-quantitive RT-PCR (P=0.0004). In concordance with the results from OSCC tissues, the relative transcript levels of MAL gene were significantly five-fold lower in 9 OSCC cell lines than in normal oral epithelium cells by real-time PCR. A significant reduction in cell proliferation and a change in cell cycle phase were observed in MAL transfectants. Matrigel assays demonstrated that the OSCC cell invasiveness was significantly decreased. A strong increase in population of apoptotic cells by MAL gene transfer was also observed. Xenografts growth was inhibited in cells expressing MAL compared with cells not expressing MAL (P=0.018).

Conclusion: The results showed that MAL is able to suppress malignant phenotypes of OSCC, such as proliferation, apoptosis and invasion, and inhibit tumorigenicity in vivo by transient and stable transfection and suggested MAL might be a candidate of tumor suppression gene. This gene may become a promising target molecule for OSCC diagnosis and gene therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4686.
Abstract 4687: Gene expression of breast cancer with CD44+CD24-/low genotype
Authors: Tsunoda Yuko , Sakamoto Masaaki , Suzuki Kenya , Sawada Terumasa , Sasaki Akiko , Yamamoto Gou , Isobe Tomohide , Tachikawa Tetsuhiko
Published: February 2011

Purpose: Breast cancer cells with a CD44+CD24-/low phenotype have been suggested to have tumor-initiating properties with stem cell-like. The purpose of this study is to clarify the gene expression profiling of cells with different CD44/CD24 genotypes within breast cancer.

Methods: Laser-captured microdissection was used to select the isolation of cancer cells in 36 frozen tissues of breast cancer, and RNA extracted from these cells was examined by real-time RT-PCR to quantify CD44 and CD24 expressions. Human stem cell RT2 profiler PCR array was used for gene expression analysis in the groups of different CD44/CD24 genotype. Associations between different CD44/CD24 genotypes and clinical variables were assessed by Fisher's exact test, and the Mann-Whitney U test was used for gene expression profiling. P<0.05 was considered significant.

Results: Thirty-six tumors were divided into 3 groups. Group A was composed of CD44+CD24-/low genotype, in which the ratio of CD44/CD24 was >1.0. Group B was composed of CD44+CD24+ genotype, in which that was 0.1 < ratio ≤ 1.0. In group C of CD44-/lowCD24+ genotype, that was < 0.1. The number of tumors in group A, B and C was 5, 29 and 2, respectively. Regarding the correlation of CD44/CD24 status with tumor characteristics, tumors of group A were significantly associated with lymph node metastasis compared with those of group B (P=0.029). The number of tumor with HER 2 score 3 within group A and B was 0 and 11, respectively. There was no significantly difference in ER status and tumor size. As far as the signaling pathways, the number of expression genes for Notch pathway in group A was significantly greater than in group B (P=0.028), and that for Wnt pathway was not significantly different between group A and B. Overexpressed gene in group A was NUMB, which is related to the programmed cell death. MYST1 and SOX2 tended to show higher levels of group A than of group B.

Conclusion: This study suggests that Notch pathway may be more important than Wnt pathway on breast cancer with CD44+CD24-/low genotype, and the CD44+CD24-/low status would be associated with low/negative HER2 expression. Moreover, the gene of self-renewal markers, such as MYST1, SOX2 and NUMB might be a target for therapy against breast cancer with low/negative HER2 expression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4687.
Abstract 4691: Evaluating the effectiveness of community based breast cancer and prostate cancer interventions among African Americans
Authors: Louis Judithe , Brown Camille , Cannon Crystal , Wilkins Jeffrey , Cameron Amura , Patrice Jasmine , Livingston Jonathan , Singleton Dorothy
Published: February 2011

Health disparities among African Americans have been of key concern for the past decade. High rates of morbidity due to HIV/AIDS, heart disease, and cancer have led to the creation of new and more innovative efforts to educate African Americans about health disparities. The current paper is an assessment of the use of Community Theater to educate African Americans about breast and prostate cancers. To assess the effectiveness of theater, a team of health professionals, psychologists, and biomedical scientists were convened to develop two scripts written by a local playwright: “Stealing Clouds” which focused on breast cancer and “Fourth and a Mile” which focused on prostate cancer. A retrospective pre/post-test design with a 3-month delayed time series follow up was employed using surveys from participants upon viewing the plays. Results of both theatre presentations indicated that the plays were significant in increasing knowledge and awareness about both breast and prostate cancers. Furthermore, results indicated that participants, as a result of viewing the plays, reported increased likelihood of engaging in healthier behaviors (i.e., physician visits, testing). To assess the impact of Community Theatre as an effective medium, data was collected from a local health center indicating higher increases in mammography and prostate cancer exams as results of the plays. Given the high rates of cancer among African Americans, continued creation and evaluation of the effectiveness of such community-based efforts should be of key import in reducing and addressing cancer disparities.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4691.
Abstract 4695: Elevated levels of biomarkers of volatile organic carcinogens and toxicants in Chinese women who regularly cook at home
Authors: Carmella Steven G. , Hecht Stephen S. , Seow Adeline , Wang Mingyao , Wang Renwei , Meng Lei , Koh Woon-Puay , Chen Menglan , Han Shaomei , Yu Mimi C. , Yuan Jian-Min
Published: February 2011

Chinese women in Asia have higher rates of lung cancer than in many other places in the world. In China, the age standardized incidence rate for female lung cancer is 19.9 per 100,000 and in Singapore it is 17.5, while rates are far lower (1.5-9.2) in countries such as Vietnam, Indonesia, India, and Russia. Cigarette smoking is not the major risk factor for lung cancer in these women, nor is secondhand tobacco smoke exposure. Some studies demonstrate the role of fumes from indoor heating under special circumstances. However, multiple epidemiologic studies document an association between lung cancer in Chinese women and Chinese-style cooking with oils heated at high temperatures. Carcinogens and toxicants have been identified in fumes generated during high temperature cooking with various oils but there have been no biomarker studies to assess carcinogen and toxicant uptake in Chinese women who regularly cook at home. We quantified urinary mercapturic acid biomarkers of exposure to the volatile compounds acrolein, crotonaldehyde, benzene, 1,3-butadiene, and ethylene oxide; and urinary metabolites of the non-volatile combustion products pyrene and phenanthrene in 54 non-smoking, non alcohol-drinking Chinese women who reported regularly doing home cooking and 50 non-smoking, non-alcohol drinking women randomly selected from the Singapore Chinese Health Study as controls. Compared with controls, women who engaged in regular home cooking had significantly higher levels of mercapturic acids of acrolein [geometric mean 1959 pmol/mg creatinine (95% CI 1554-2467) vs.1370 (95% CI 1077-1742), P = 0.038], crotonaldehyde [geometric mean 231.6 pmol/mg creatinine (95% CI 193-277) vs. 142.2 (95% CI 118-171) P = 0.0004], and benzene [geometric mean 0.58 pmol/mg creatinine (95% CI 0.44-0.78) vs. 0.18 (95% CI 0.14-0.24) P < 0.0001]. No significant differences were found in levels of mercapturic acids of 1,3-butadiene or metabolites of pyrene and phenanthrene between the two groups whereas levels of the ethylene oxide mercapturic acid were significantly higher in the controls than in the cooking group. The higher levels of the mercapturic acid of benzene, a multi-organ carcinogen, in the women who cooked, are particularly notable. The results of this study showing increased exposure to the volatile toxicants and carcinogens acrolein, crotonaldehyde, and benzene in Chinese women who regularly cook provide a plausible lead for further investigating the role of volatile compounds generated during cooking as causes of lung cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4695.
Abstract 4697: Liquid chromatography-electrospray ionization tandem mass spectrometry analysis of 7-ethylguanine in human urine
Authors: Balbo Silvia , Hochalter J. Bradley , Villalta Peter W. , Carmella Steven G. , Hecht Stephen S.
Published: February 2011

Considerable evidence in the literature demonstrates the exposure of humans to an unknown ethylating agent. Previous studies have demonstrated the presence of 3-ethyl-Ade in urine, 7-ethylguanine (7-ethyl-Gua) in urine and human liver DNA, 7-ethyldGuo and O4-ethyl-dThd in human lung DNA, and ethylvaline in hemoglobin. Some studies also report higher levels of ethyl adducts in smokers compared to nonsmokers, and there is convincing evidence for an uncharacterized ethylating agent in cigarette smoke. To further investigate this question and evaluate the potential role of ethylating agents in smoking-induced cancer we have developed a liquid chromatography-electrospray ionization tandem mass spectrometry-selected reaction monitoring (LC-ESI-MS/MS-SRM) method for analysis of 7-ethyl-Gua in human urine.

A two-dimensional chromatography approach was used with [15N5]7-ethyl-Gua as the internal standard. Human urine was diluted with 15 mM ammonium acetate and partially purified by a solid-phase extraction cartridge (Strata-X 33 µm, 30 mg/1 mL, Phenomenex) and a mixed mode cation exchange reversed-phase extraction cartridge (Oasis MCX, 30 mg/cartridge, Waters). The fraction from this last purification step containing the analyte (30% MeOH, 5% NH4OH) was dried and redissolved in 80% acetonitrile, 0.1% formic acid and purified by HPLC on a YMC Polyamino II column (150 × 1.0 mm I.D. 5 μm). The fraction containing 7-ethyl-Gua was collected, dried, dissolved in H2O and analysed by LC-ESI-MS/MS-SRM using a Thermo Hypersil Gold PFP column (150 × 0.5 mm I.D. 3 μm). The analysis was carried out using the transition m/z 180 [M + H]+ → m/z 152 [Gua+ H]+ for 7-ethyl-Gua and m/z 185 → m/z 157 for the internal standard. Recovery was about 50%. The detection limit was approximately 10 fmol on column. The peaks attributed to 7-ethyl-Gua and the labeled internal standard had identical retention time and were observed in most of the human urine samples analyzed. The identity of the peak was confirmed by re-injecting the samples after spiking with a standard solution of 7-ethyl-Gua. Twenty-one human urine samples (1 mL) from 11 smokers and 10 nonsmokers were analyzed. 7-Ethyl-Gua was clearly detected in 8 out of 11 samples from smokers and 5 out of 10 samples from nonsmokers. These results strongly indicate that 7-ethyl-Gua is present in human urine and demonstrate that it can be detected and quantified. Further investigation is required to clarify its relationship to cigarette smoking.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4697.
Abstract 47: Pyruvate kinase M2 is a target of the tumor-suppressive microRNA-326 and regulates the survival of glioma cells
Authors: Kefas Benjamin A.
Published: February 2011

Emerging studies have identified microRNAs as possible therapeutic tools for the treatment of glioma, the most aggressive brain tumor. Their important targets in this tumor are not well understood. We recently found that the Notch pathway is a target of microRNA-326. Ectopic expression of microRNA-326 in glioma and glioma stem cells induced their apoptosis and reduced their metabolic activity. Computational target gene prediction revealed pyruvate kinase type M2 (PKM2) as another target of microRNA-326. PKM2 has recently been shown to play a key role in cancer cell metabolism. To investigate whether it might be a functionally important target of miR-326, we used RNA interference to knock down PKM2 expression in glioma cells. Transfection of established glioma and glioma stem cells with PKM2 siRNA reduced their growth, cellular invasion, metabolic activity with decreased ATP and glutathione levels, and activated AMP-activated protein kinase. The cytotoxic effects exhibited by PKM2 knockdown in glioma and glioma stem cells were not observed in transformed human astrocytes. Western blot analysis of human glioblastoma specimens showed high levels of PKM2 protein but none was observed in normal brain samples. Our data suggest PKM2 inhibition as a therapy for glioblastoma, with the potential for minimal toxicity to the brain.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 47.
Abstract 4701: DNA-adducts levels and chromosomal aberrations in relation to smoking history and topography in smokers
Authors: Dumitrescu Ramona G. , Zheng Yun-Ling , Yang Yang , Seillier-Moiseiwitsch Francoise , Spernak Stephanie M. , Loffredo Christopher , Phillips David H. , Shields Peter G.
Published: February 2011

Introduction: Tobacco smoking is the major cause of morbidity and mortality in developed countries. Important determinants of tobacco carcinogen exposure include the smoking history and smoking topography. Tobacco carcinogens-induced DNA adducts and chromosomal aberrations have been associated with cancer development. It is unknown if the choice of cigarettes and smoking topography lead to differences in carcinogen-DNA adduct levels and chromosomal aberrations.

Methods: We studied 207 population-based smokers (97 Caucasians and 110 African Americans). The subjects smoked two cigarettes, one hour apart, and the smoking topography was measured. We assessed three parameters of smoke exposure (serum cotinine, nicotine boost and carbon monoxide boost) in order to collect corroborative evidence that reflect different types of exposure. Carcinogen-DNA adducts and chromosomal aberrations in lymphocytes were determined by using the 32P-postlabeling analysis and standard cytogenetic method, respectively.

Results: The chance of having high level of DNA-adducts was significantly higher in males than females (OR: 1.9; 95%CI: 1.09-3.31). Weakly associations of the DNA-adducts were observed with age and race (higher in older people and African-Americans smokers) (OR: 1.7; 95%CI: 0.97-2.90; OR: 1.5; 95%CI: 0.87-2.60). DNA-adducts were significantly associated with the baseline and post cigarette CO level for the first (p-value=0.001; 0.007 respectively) and second cigarette (p-value=<0.001; 0.001 respectively), with the total number of puffs for the second cigarette (p-value=0.04) and with the cumulative topography (p-value=0.03). Marginal associations were observed for the changes in CO2 pre and post cigarettes (p-value=0.08), the average inter-puff interval and the total puff volume for the second cigarette (p-value=0.06; 0.07 respectively). The changes in CO2 pre and post first and second cigarette were significantly associated with the mean number of chromosomal aberrations per cell (p-value=0.05; 0.01 respectively).

Conclusion: In this study we observed potentially important associations of smoking topography with both DNA adduct levels and chromosomal aberrations. Larger study sets will be necessary to clarify the magnitude of these associations and to account for baseline differences related to gender, age, and race. Such an approach could provide useful information for understanding differential susceptibility to lung cancer among smokers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4701.