bluuebunny/crossref_metadata_2025 huggingface dataset:
Supplementary Figure from Carbon Monoxide Activates PERK-Regulated Autophagy to Induce Immunometabolic Reprogramming and Boost Antitumor T-cell Function
Authors: Chakraborty Paramita , Parikh Rasesh Y. , Choi Seungho , Tran Danh , Gooz Monika , Hedley Zachariah T. , Kim Do-Sung , Pytel Dariusz , Kang Inhong , Nadig Satish N. , Beeson Gyda C. , Ball Lauren , Mehrotra Meenal , Wang Hongjun , Berto Stefano , Palanisamy Viswanathan , Li Hong , Chatterjee Shilpak , Rodriguez Paulo C. , Maldonado Eduardo N. , Diehl J. Alan , Gangaraju Vamsi K. , Mehrotra Shikhar
Published: March 2023

Supplementary Figure from Carbon Monoxide Activates PERK-Regulated Autophagy to Induce Immunometabolic Reprogramming and Boost Antitumor T-cell Function
Supplementary Data from CDK4/6 Inhibition Enhances Oncolytic Virus Efficacy by Potentiating Tumor-Selective Cell Killing and T-cell Activation in Refractory Glioblastoma
Authors: Xiao Jingshu , Liang Jiaming , Fan Junjie , Hou Panpan , Li Xiaodong , Zhang Haipeng , Li Kai , Bu Lang , Li Ping , He Miao , Zhong Yongheng , Guo Liping , Jia Penghui , Xiao Qiaoqiao , Wu Junyu , Peng Hong , Li Chunmei , Xing Fan , Guo Deyin
Published: March 2023

Supplementary Data from CDK4/6 Inhibition Enhances Oncolytic Virus Efficacy by Potentiating Tumor-Selective Cell Killing and T-cell Activation in Refractory Glioblastoma
Supplementary Figure 6 from Monounsaturated and Diunsaturated Fatty Acids Sensitize Cervical Cancer to Radiation Therapy
Authors: Muhammad Naoshad , Ruiz Fiona , Stanley Jennifer , Rashmi Ramachandran , Cho Kevin , Jayachandran Kay , Zahner Michael C. , Huang Yi , Zhang Jin , Markovina Stephanie , Patti Gary J. , Schwarz Julie K.
Published: March 2023

Oleic acid and radiation alter p53 related gene expression
Table S20 from Integrative Single-Cell Analysis Reveals Transcriptional and Epigenetic Regulatory Features of Clear Cell Renal Cell Carcinoma
Authors: Yu Zhenyuan , Lv Yufang , Su Cheng , Lu Wenhao , Zhang RuiRui , Li Jiaping , Guo Bingqian , Yan Haibiao , Liu Deyun , Yang Zhanbin , Mi Hua , Mo Linjian , Guo Yi , Feng Wenyu , Xu Haotian , Peng Wenyi , Cheng Jiwen , Nan Aruo , Mo Zengnan
Published: March 2023

qPCR results after ASO (5717 and 5608) treatment of 2 lncRNAs in caki-2 and 786-O cell lines.Three or six PCR by-wells were prepared for each sample. NC, negative control.
Supplementary MovieS 1a from Immunotherapeutic Targeting and PET Imaging of DLL3 in Small-Cell Neuroendocrine Prostate Cancer
Authors: Chou Jonathan , Egusa Emily A. , Wang Sinan , Badura Michelle L. , Lee Fei , Bidkar Anil P. , Zhu Jun , Shenoy Tanushree , Trepka Kai , Robinson Troy M. , Steri Veronica , Huang Jiaoti , Wang Yuzhuo , Small Eric J. , Chan Emily , Stohr Bradley A. , Ashworth Alan , Delafontaine Brant , Rottey Sylvie , Cooke Keegan S. , Hashemi Sadraei Nooshin , Yu Brian , Salvati Mark , Bailis Julie M. , Feng Felix Y. , Flavell Robert R. , Aggarwal Rahul
Published: March 2023

Supplemental Movies 1a-1d: 22Rv1-LacZ and 22Rv1-DLL3 cells co-cultures. Time-lapse movies of 22Rv1-LacZ and 22Rv1-DLL3 cells (labeled with NuclightRed) co-cultured with T cells and either NT control BiTE® or AMG 757. T cells were plated at an E:T ratio of 5:1. Images were captured every 6 hours at 10x magnification.
a. 22Rv1-LacZ cells and T cells treated with NT control BiTE®.
b. 22Rv1-DLL3 cells and T cells treated with NT control BiTE®.
c. 22Rv1-LacZ cells and T cells treated with AMG 757.
d. 22Rv1-DLL3 cells and T cells treated with AMG 757.
FigureS8 from Immunotherapeutic Targeting and PET Imaging of DLL3 in Small-Cell Neuroendocrine Prostate Cancer
Authors: Chou Jonathan , Egusa Emily A. , Wang Sinan , Badura Michelle L. , Lee Fei , Bidkar Anil P. , Zhu Jun , Shenoy Tanushree , Trepka Kai , Robinson Troy M. , Steri Veronica , Huang Jiaoti , Wang Yuzhuo , Small Eric J. , Chan Emily , Stohr Bradley A. , Ashworth Alan , Delafontaine Brant , Rottey Sylvie , Cooke Keegan S. , Hashemi Sadraei Nooshin , Yu Brian , Salvati Mark , Bailis Julie M. , Feng Felix Y. , Flavell Robert R. , Aggarwal Rahul
Published: March 2023

FigureS8
FigureS6 from Immunotherapeutic Targeting and PET Imaging of DLL3 in Small-Cell Neuroendocrine Prostate Cancer
Authors: Chou Jonathan , Egusa Emily A. , Wang Sinan , Badura Michelle L. , Lee Fei , Bidkar Anil P. , Zhu Jun , Shenoy Tanushree , Trepka Kai , Robinson Troy M. , Steri Veronica , Huang Jiaoti , Wang Yuzhuo , Small Eric J. , Chan Emily , Stohr Bradley A. , Ashworth Alan , Delafontaine Brant , Rottey Sylvie , Cooke Keegan S. , Hashemi Sadraei Nooshin , Yu Brian , Salvati Mark , Bailis Julie M. , Feng Felix Y. , Flavell Robert R. , Aggarwal Rahul
Published: March 2023

FigureS6
FigureS2 from Immunotherapeutic Targeting and PET Imaging of DLL3 in Small-Cell Neuroendocrine Prostate Cancer
Authors: Chou Jonathan , Egusa Emily A. , Wang Sinan , Badura Michelle L. , Lee Fei , Bidkar Anil P. , Zhu Jun , Shenoy Tanushree , Trepka Kai , Robinson Troy M. , Steri Veronica , Huang Jiaoti , Wang Yuzhuo , Small Eric J. , Chan Emily , Stohr Bradley A. , Ashworth Alan , Delafontaine Brant , Rottey Sylvie , Cooke Keegan S. , Hashemi Sadraei Nooshin , Yu Brian , Salvati Mark , Bailis Julie M. , Feng Felix Y. , Flavell Robert R. , Aggarwal Rahul
Published: March 2023

FigureS2
Supplementary video from Phagocytosis of Glioma Cells Enhances the Immunosuppressive Phenotype of Bone Marrow–Derived Macrophages
Authors: Wu Min , Wu Lingxiang , Wu Wei , Zhu Mengyan , Li Jianyu , Wang Ziyu , Li Jie , Ding Rong , Liang Yuan , Li Liangyu , Zhang Tingting , Huang Bin , Cai Yun , Li Kening , Li Lu , Zhang Rui , Hu Baoli , Lin Fan , Wang Xiuxing , Zheng Siyuan , Chen Jian , You Yongping , Jiang Tao , Zhang Junxia , Chen Hongshan , Wang Qianghu
Published: March 2023

Macrophages phagocytose tumor cells.
Supplementary Data from HAND2 Assists MYCN Enhancer Invasion to Regulate a Noradrenergic Neuroblastoma Phenotype
Authors: Xu Man , Sun Ming , Zhang Xiyuan , Nguyen Rosa , Lei Haiyan , Shern Jack F. , Thiele Carol J. , Liu Zhihui
Published: March 2023

Table S8 shows genes altered after silencing both HAND2 and MYCN.
Supplemental Figure 2 from NFE2L2 Mutations Enhance Radioresistance in Head and Neck Cancer by Modulating Intratumoral Myeloid Cells
Authors: Guan Li , Nambiar Dhanya K. , Cao Hongbin , Viswanathan Vignesh , Kwok Shirley , Hui Angela B. , Hou Yuan , Hildebrand Rachel , von Eyben Rie , Holmes Brittany J. , Zhao Junfei , Kong Christina S. , Wamsley Nathan , Zhang Weiruo , Major Michael B. , Seol Seung W. , Sunwoo John B. , Hayes D. Neil , Diehn Maximilian , Le Quynh-Thu
Published: March 2023

The relationship between NFE2L2 mutation and radiation resistance in OC cancer. Kaplan-Meier analysis of PFS and OSin OC patients treated with surgery and postoperative (chemo)radiotherapy.
Figure S1 from Reimagining Cancer: Moving from the Cellular to the Tissue Level
Authors: Demicheli Romano , Hrushesky William J.M.
Published: March 2023

Self-organizing process
Data from Mitochondrially Targeted p53 Has Tumor Suppressor Activities In vivo
Authors: Talos Flaminia , Petrenko Oleksi , Mena Patricio , Moll Ute M.
Published: March 2023

Abstract
Complex proapoptotic functions are essential for the tumor suppressor activity of p53. We recently described a novel transcription-independent mechanism that involves a rapid proapoptotic action of p53 at the mitochondria and executes the shortest known circuitry of p53 death signaling. Here, we examine if this p53-dependent mitochondrial program could be exploited for tumor suppression in vivo. To test this, we engage Eμ-Myc transgenic mice, a well-established model of p53-dependent lymphomagenesis. We show that exclusive delivery of p53 to the outer mitochondrial membrane confers a significant growth disadvantage on Eμ-Myc–transformed B-cells of p53-deficient or alternate reading frame–deficient genotypes, resulting in efficient induction of apoptosis and impinged proliferation. Conversely, normal cells from thymus, spleen, and bone marrow showed poor infectivity with these viruses. This proof-of-principle experiment shows that exclusive reliance on the direct mitochondrial program exerts a significant tumor suppressor activity in vivo. Our in vivo data on the direct mitochondrial apoptotic p53 program lays the groundwork to further investigate its efficacy and safety and to address its possible therapeutic value in the future.
Data from Different S/M Checkpoint Responses of Tumor and Non–Tumor Cell Lines to DNA Replication Inhibition
Authors: Rodríguez-Bravo Verónica , Guaita-Esteruelas Sandra , Salvador Noelia , Bachs Oriol , Agell Neus
Published: March 2023

Abstract
Cell cycle checkpoint abrogation, especially the inhibition of Chk1 in combination with DNA-damaging treatments, has been proposed as a promising way of sensitizing cancer cells. However, less is known about the possibility to selectively affect tumor cells when they are treated with agents that block DNA synthesis in combination with replication checkpoint inhibitors. Here, we present clear insights in the different responses of tumor and non-transformed cells to the inhibition of DNA replication with hydroxyurea in combination with checkpoint abrogation via inhibition of Ataxia telangiectasia–mutated– (ATM) and Rad3-related/ATM (ATR/ATM) and Chk1 kinases. Interestingly, we find that non-transformed cell lines activate ATR/ATM- and Chk1-independent pathways in response to replication inhibition to prevent mitotic entry with unreplicated DNA. In contrast, tumor cell lines such as HCT116 and HeLa cells rely entirely on Chk1 activity for a proper response to replication inhibitors. Our results show that p38 is activated in response to hydroxyurea treatment and collaborates with Chk1 to prevent mitotic entry in non-transformed cell lines by maintaining cyclin B1/Cdk1 complexes inactive. Furthermore, DNA replication arrest down-regulates cyclin B1 promoter activity in non-transformed cells, but not in tumor cells in a Chk1- and p38-independent way. Thus, our data show that non-transformed cells present a more robust DNA replication checkpoint response compared with tumor cells that involves activation of the p38 pathway. We show that some of these responses to replication block can be lost in tumor cells, causing a defective checkpoint and providing a rationale for tumor-selective effects of combined therapies. [Cancer Res 2007;67(24):11648–56]
Data from Differing Src Signaling Levels Have Distinct Outcomes in Drosophila
Authors: Vidal Marcos , Warner Stephen , Read Renee , Cagan Ross L.
Published: March 2023

Abstract
High levels of Src activity are found in a broad spectrum of cancers. The roles of Src and its negative regulator Csk have been extensively studied, although results have often proved contradictory or the relevance to whole organisms is unclear. In Drosophila, overexpression of either Src orthologue resulted in apoptotic cell death, but paradoxically, reducing dCsk activity led to over-proliferation and tissue overgrowth. Here, we show that in Drosophila epithelia in situ, the levels of Src signaling determine the cellular outcome of Src activation. Apoptotic cell death was triggered specifically at high Src signaling levels; lower levels directed antiapoptotic signals while promoting proliferation. Furthermore, our data indicate that expression of kinase-dead Src isoforms do not necessarily act as dominant-negative factors, but can instead increase Src pathway activity, most likely by titrating Csk activity away from endogenous Src. The importance of Src activity levels was emphasized when we examined oncogenic cooperation between Src and Ras: malignant overgrowth was observed specifically when high Src signaling levels were achieved. We propose a model in which low levels of Src signaling promote survival and proliferation during early stages of tumorigenesis, whereas strong Src signaling, coupled with antiapoptotic signals, directs invasive migration and metastasis during advanced tumor stages. [Cancer Res 2007;67(21):10278–85]
Data from A Novel Bone Morphogenetic Protein Signaling in Heterotypic Cell Interactions in Prostate Cancer
Authors: Yang Shangxin , Pham Linda K. , Liao Chun-Peng , Frenkel Baruch , Reddi A. Hari , Roy-Burman Pradip
Published: March 2023

Abstract
We examined the effect of the extracellular bone morphogenetic protein (BMP) 2 and 7, which are up-regulated in the prostate adenocarcinomas of the conditional Pten deletion mouse model, on primary cultures of cancer-associated fibroblasts (CAF) derived from these tumors. In the CAF, we show that BMP2 or BMP7, but not transforming growth factor β-1, can strikingly stimulate secretion of stromal cell–derived factor-1 (SDF-1), also known as CXCL12. The CAF cells express type I and type II BMP receptors as well as the receptor for SDF-1, CXCR4. SDF-1 activation is associated with BMP-induced Smad phosphorylation, and the stimulatory effect is blocked by BMP antagonist, noggin. The findings that BMP treatment can increase SDF-1 pre-mRNA levels in a time-dependent manner and actinomycin D treatment can abolish stimulatory effect of BMP suggest a transcriptional modulation of SDF-1 by BMP signaling. Using a human microvascular endothelial cell line, we show that SDF-1 present in the conditioned medium from the stimulated CAF can significantly induce tube formation, an effect relating to angiogenic function. Furthermore, we found that BMP2 can also protect the CAF from serum starvation–induced apoptosis independent of SDF-1, implying that BMP may induce other factors to sustain the survival of these cells. In short, this report establishes a novel BMP-SDF-1 axis in the prostate tumor along with a new prosurvival effect of BMP that when considered together with our previously described oncogenic properties of BMP indicate a circuitry for heterotypic cell interactions potentially critical in prostate cancer. [Cancer Res 2008;68(1):198–205]
Data from Syndecan-2 Affects the Basal and Chemotherapy-Induced Apoptosis in Osteosarcoma
Authors: Orosco Armelle , Fromigué Olivia , Bazille Céline , Entz-Werle Natacha , Levillain Pierre , Marie Pierre J. , Modrowski Dominique
Published: March 2023

Abstract
Syndecans are transmembrane heparan sulfate proteoglycans controlling cell adhesion, migration, and proliferation. We previously showed that syndecan-2 is involved in the control of apoptosis in cultured osteosarcoma cells. These data led us to the hypothesis that syndecan-2 may play a role in the apoptotic signaling in bone tumors. We immunohistochemically analyzed tissue sections from biopsies from 21 patients with well-characterized osteosarcoma. These tissues expressed low levels of syndecan-2 compared with osteoblasts and osteocytes in normal bone. Cultured human osteosarcoma cells also produced lower mRNA levels of syndecan-2 than normal osteoblastic cells. Moreover, the presence of syndecan-2 correlated with spontaneous apoptosis in osteosarcoma tissues as assessed by detection of DNA fragmentation in situ. Overexpression of syndecan-2 resulted in decreased number of migrating and invading U2OS osteosarcoma cells in Matrigel. In addition, overexpression of syndecan-2 sensitized human osteosarcoma cells to chemotherapy-induced apoptosis, increasing the response to methotrexate, doxorubicin, and cisplatin. Consistently, knockdown of the proteoglycan using stable transfection with a plasmid coding small interfering RNA resulted in inhibition of chemotherapy-induced apoptosis. Analysis of syndecan-2 expression both in biopsies and in corresponding postchemotherapy-resected tumors, as well as in cells treated with methotrexate or doxorubicin, showed that the cytotoxic action of chemotherapy can be associated with an increase in syndecan-2. These results provide support for a tumor-suppressor function for syndecan-2 and suggest that dysregulation of apoptosis may be related to abnormal syndecan-2 expression or induction in osteosarcoma. Moreover, our data identify syndecan-2 as a new factor mediating the antioncogenic effect of chemotherapeutic drugs. [Cancer Res 2007;67(8):3708–14]
Data from FBXW7/hCDC4 Is a General Tumor Suppressor in Human Cancer
Authors: Akhoondi Shahab , Sun Dahui , von der Lehr Natalie , Apostolidou Sophia , Klotz Kathleen , Maljukova Alena , Cepeda Diana , Fiegl Heidi , Dofou Dimitra , Marth Christian , Mueller-Holzner Elisabeth , Corcoran Martin , Dagnell Markus , Nejad Sepideh Zabihi , Nayer Babak Noori , Zali Mohammad Reza , Hansson Johan , Egyhazi Susanne , Petersson Fredrik , Sangfelt Per , Nordgren Hans , Grander Dan , Reed Steven I. , Widschwendter Martin , Sangfelt Olle , Spruck Charles
Published: March 2023

Abstract
The ubiquitin-proteasome system is a major regulatory pathway of protein degradation and plays an important role in cellular division. Fbxw7 (or hCdc4), a member of the F-box family of proteins, which are substrate recognition components of the multisubunit ubiquitin ligase SCF (Skp1-Cdc53/Cullin-F-box-protein), has been shown to mediate the ubiquitin-dependent proteolysis of several oncoproteins including cyclin E1, c-Myc, c-Jun, and Notch. The oncogenic potential of Fbxw7 substrates, frequent allelic loss in human cancers, and demonstration that mutation of FBXW7 cooperates with p53 in mouse tumorigenesis have suggested that Fbxw7 could function as a tumor suppressor in human cancer. Here, we carry out an extensive genetic screen of primary tumors to evaluate the role of FBXW7 as a tumor suppressor in human tumorigenesis. Our results indicate that FBXW7 is inactivated by mutation in diverse human cancer types with an overall mutation frequency of ∼6%. The highest mutation frequencies were found in tumors of the bile duct (cholangiocarcinomas, 35%), blood (T-cell acute lymphocytic leukemia, 31%), endometrium (9%), colon (9%), and stomach (6%). Approximately 43% of all mutations occur at two mutational “hotspots,” which alter Arg residues (Arg465 and Arg479) that are critical for substrate recognition. Furthermore, we show that Fbxw7Arg465 hotspot mutant can abrogate wild-type Fbxw7 function through a dominant negative mechanism. Our study is the first comprehensive screen of FBXW7 mutations in various human malignancies and shows that FBXW7 is a general tumor suppressor in human cancer. [Cancer Res 2007;67(19):9006–12]
Data from Medulloblastomas Derived from Cxcr6 Mutant Mice Respond to Treatment with a Smoothened Inhibitor
Authors: Sasai Ken , Romer Justyna T. , Kimura Hiromichi , Eberhart Derek E. , Rice Dennis S. , Curran Tom
Published: March 2023

Abstract
The sonic hedgehog (Shh) pathway is activated in ∼30% of human medulloblastoma resulting in increased expression of downstream target genes. In about half of these cases, this has been shown to be a consequence of mutations in regulatory genes within the pathway, including Ptc1, Smo, and Sufu. However, for some tumors, no mutations have been detected in known pathway genes. This suggests that either mutations in other genes promote tumorigenesis or that epigenetic alterations increase pathway activity in these tumors. Here, we report that 3% to 4% of mice lacking either one or both functional copies of Cxcr6 develop medulloblastoma. Although CXCR6 is not known to be involved in Shh signaling, tumors derived from Cxcr6 mutant mice expressed Shh pathway target genes including Gli1, Gli2, Ptc2, and Sfrp1, indicating elevated pathway activity. Interestingly, the level of Ptc1 expression was decreased in tumor cells although two normal copies of Ptc1 were retained. This implies that reduced CXCR6 function leads to suppression of Ptc1 thereby increasing Smoothened function and promoting tumorigenesis. We used a direct transplant model to test the sensitivity of medulloblastoma arising in Cxcr6 mutant mice to a small-molecule inhibitor of Smoothened (HhAntag). We found that transplanted tumors were dramatically inhibited in mice treated for only 4 days with HhAntag. These findings suggest that HhAntag may be effective against tumors lacking mutations in known Shh pathway genes. [Cancer Res 2007;67(8):3871–7]
Data from Placental Growth Factor-1 Attenuates Vascular Endothelial Growth Factor-A–Dependent Tumor Angiogenesis during β Cell Carcinogenesis
Authors: Schomber Tibor , Kopfstein Lucie , Djonov Valentin , Albrecht Imke , Baeriswyl Vanessa , Strittmatter Karin , Christofori Gerhard
Published: March 2023

Abstract
Members of the vascular endothelial growth factor (VEGF) family are critical players in angiogenesis and lymphangiogenesis. Although VEGF-A has been shown to exert fundamental functions in physiologic and pathologic angiogenesis, the exact role of the VEGF family member placental growth factor (PlGF) in tumor angiogenesis has remained controversial. To gain insight into PlGF function during tumor angiogenesis, we have generated transgenic mouse lines expressing human PlGF-1 in the β cells of the pancreatic islets of Langerhans (Rip1PlGF-1). In single-transgenic Rip1PlGF-1 mice, intra-insular blood vessels are found highly dilated, whereas islet physiology is unaffected. Upon crossing of these mice with the Rip1Tag2 transgenic mouse model of pancreatic β cell carcinogenesis, tumors of double-transgenic Rip1Tag2;Rip1PlGF-1 mice display reduced growth due to attenuated tumor angiogenesis. The coexpression of transgenic PlGF-1 and endogenous VEGF-A in the β tumor cells of double-transgenic animals causes the formation of low-angiogenic hPlGF-1/mVEGF-A heterodimers at the expense of highly angiogenic mVEGF-A homodimers resulting in diminished tumor angiogenesis and reduced tumor infiltration by neutrophils, known to contribute to the angiogenic switch in Rip1Tag2 mice. The results indicate that the ratio between the expression levels of two members of the VEGF family of angiogenic factors, PlGF-1 and VEGF-A, determines the overall angiogenic activity and, thus, the extent of tumor angiogenesis and tumor growth. [Cancer Res 2007;67(22):10840–8]
Data from Arsenic Trioxide Sensitizes Human Glioma Cells, but not Normal Astrocytes, to TRAIL-Induced Apoptosis via CCAAT/Enhancer-Binding Protein Homologous Protein–Dependent DR5 Up-regulation
Authors: Kim Eun Hee , Yoon Mi Jin , Kim Seung U. , Kwon Taeg Kyu , Sohn Seonghyang , Choi Kyeong Sook
Published: March 2023

Abstract
The current study shows that treatment of tumor necrosis factor–related apoptosis-inducing ligand (TRAIL)–resistant glioma cells with a combination of TRAIL and subtoxic doses of arsenic trioxide (As2O3) induces rapid apoptosis. Whereas TRAIL-mediated proteolytic processing of procaspase-3 was partially blocked in glioma cells, treatment with As2O3 efficiently recovered TRAIL-induced activation of caspases. We also found that As2O3 treatment of glioma cells significantly up-regulated DR5, a death receptor of TRAIL. Furthermore, suppression of DR5 expression by small interfering RNA (siRNA) inhibited As2O3/TRAIL-induced apoptosis of U87MG glioma cells, suggesting that DR5 up-regulation is critical for As2O3-induced sensitization of glioma cells to TRAIL-mediated apoptosis. Our results also indicate that an increase in CCAAT/enhancer binding protein homologous protein (CHOP) protein levels precedes As2O3-induced DR5 up-regulation. The involvement of CHOP in this process was confirmed by siRNA-mediated CHOP suppression, which not only attenuated As2O3-induced DR5 up-regulation but also inhibited the As2O3-stimulated TRAIL-induced apoptosis. These results therefore suggest that the CHOP-mediated DR5 up-regulation, brought about by As2O3, stimulates the TRAIL-mediated signaling pathway. This in turn leads to complete proteolytic processing of caspase-3, which is partially primed by TRAIL in glioma cells. In contrast to human glioma cells, astrocytes were very resistant to the combined administration of As2O3 and TRAIL, demonstrating the safety of this treatment. In addition, As2O3-mediated up-regulation of CHOP and DR5, as well as partial proteolytic processing of procaspase-3 by TRAIL, was not induced in astrocytes. Taken together, the present results suggest that the combined treatment of glioma cells with As2O3 plus TRAIL may provide an effective and selective therapeutic strategy. [Cancer Res 2008;68(1):266–75]
Data from P21-Activated Protein Kinase Is Overexpressed in Hepatocellular Carcinoma and Enhances Cancer Metastasis Involving c-Jun NH2-Terminal Kinase Activation and Paxillin Phosphorylation
Authors: Ching Yick-Pang , Leong Veronica Y.L. , Lee Man-Fong , Xu Hai-Tao , Jin Dong-Yan , Ng Irene Oi-Lin
Published: March 2023

Abstract
Hepatocellular carcinoma (HCC) is one of the major malignancies in the world. The prognosis of HCC is poor, due to frequent intrahepatic metastasis and tumor recurrence. P21-activated protein kinase (Pak1), a main downstream effector of small Rho GTPases, Rac1 and Cdc42, plays an important role in the regulation of cell morphogenesis, motility, mitosis, and angiogenesis. Here, we show that Pak1 gene was overexpressed in human HCCs. Overexpression of Pak1 in human HCCs was associated with more aggressive tumor behavior in terms of more metastatic phenotype and more advanced tumor stages. In addition, HCC cell line stably expressing Pak1 displayed increased cell motility rates and, conversely, knockdown of endogenous Pak1 expression by small interfering RNA reduced the migration rates of HCC cells. In an established metastatic HCC cell line, we found that Pak1 was overexpressed compared with its primary HCC cell line and this overexpression was associated with higher cell motility. Importantly, we found that c-Jun NH2-terminal kinase (JNK) was activated in HCC cell lines overexpressing Pak1. Inhibition of the JNK activity by chemical inhibitor significantly reduced the migration rates of HCC cells via attenuation of paxillin phosphorylation at Ser178. In conclusion, our results document that Pak1 is overexpressed in HCCs and plays an important role in the metastasis of HCC. The mechanism by which Pak1 induces cancer metastasis may involve activation of JNK and phosphorylation of paxillin. [Cancer Res 2007;67(8):3601–8]
Data from RET/PTC Rearrangements Preferentially Occurred in Papillary Thyroid Cancer among Atomic Bomb Survivors Exposed to High Radiation Dose
Authors: Hamatani Kiyohiro , Eguchi Hidetaka , Ito Reiko , Mukai Mayumi , Takahashi Keiko , Taga Masataka , Imai Kazue , Cologne John , Soda Midori , Arihiro Koji , Fujihara Megumu , Abe Kuniko , Hayashi Tomayoshi , Nakashima Masahiro , Sekine Ichiro , Yasui Wataru , Hayashi Yuzo , Nakachi Kei
Published: March 2023

Abstract
A major early event in papillary thyroid carcinogenesis is constitutive activation of the mitogen-activated protein kinase signaling pathway caused by alterations of a single gene, typically rearrangements of the RET and NTRK1 genes or point mutations in the BRAF and RAS genes. In childhood papillary thyroid cancer, regardless of history of radiation exposure, RET/PTC rearrangements are a major event. Conversely, in adult-onset papillary thyroid cancer among the general population, the most common molecular event is BRAFV600E point mutation, not RET/PTC rearrangements. To clarify which gene alteration, chromosome aberration, or point mutation preferentially occurs in radiation-associated adult-onset papillary thyroid cancer, we have performed molecular analyses on RET/PTC rearrangements and BRAFV600E mutation in 71 papillary thyroid cancer cases among atomic bomb survivors (including 21 cases not exposed to atomic bomb radiation), in relation to radiation dose as well as time elapsed since atomic bomb radiation exposure. RET/PTC rearrangements showed significantly increased frequency with increased radiation dose (Ptrend = 0.002). In contrast, BRAFV600E mutation was less frequent in cases exposed to higher radiation dose (Ptrend < 0.001). Papillary thyroid cancer subjects harboring RET/PTC rearrangements developed this cancer earlier than did cases with BRAFV600E mutation (P = 0.03). These findings were confirmed by multivariate logistic regression analysis. These results suggest that RET/PTC rearrangements play an important role in radiation-associated thyroid carcinogenesis. [Cancer Res 2008;68(17):7176–82]
Data from Coordinated Regulation of Pathways for Enhanced Cell Motility and Chemotaxis Is Conserved in Rat and Mouse Mammary Tumors
Authors: Wang Weigang , Wyckoff Jeffrey B. , Goswami Sumanta , Wang Yarong , Sidani Mazen , Segall Jeffrey E. , Condeelis John S.
Published: March 2023

Abstract
Correlating tumor cell behavior in vivo with patterns of gene expression has led to new insights into the microenvironment of tumor cells in the primary tumor. Until now, these studies have been done with cell line–derived tumors. In the current study, we have analyzed, in polyoma middle T oncogene (PyMT)–derived mammary tumors, tumor cell behavior and gene expression patterns of the invasive subpopulation of tumor cells by multiphoton-based intravital imaging and microarray-based expression profiling, respectively. Our results indicate that the patterns of cell behavior that contribute to invasion and metastasis in the PyMT tumor are similar to those seen previously in rat MTLn3 cell line–derived mammary tumors. The invasive tumor cells collected from PyMT mouse mammary tumors, like their counterparts from rat xenograft mammary tumors, are a population that is relatively nondividing and nonapoptotic but chemotherapy resistant and chemotactic. Changes in the expression of genes that occur uniquely in the invasive subpopulation of tumor cells in the PyMT mammary tumors that fall on the Arp2/3 complex, capping protein and cofilin pathways show a pattern like that seen previously in invasive tumor cells from the MTLn3 cell line–derived tumors. These changes predict an enhanced activity of the cofilin pathway, and this was confirmed in isolated invasive PyMT tumor cells. We conclude that changes in gene expression and their related changes in cell behavior, which were identified in the invasive tumor cells of cell line–derived tumors, are conserved in the invasive tumor cells of PyMT-derived mouse mammary tumors, although these tumor types have different genetic origins. [Cancer Res 2007;67(8):1–6]
Data from Multiple Myeloma Cell Killing by Depletion of the MET Receptor Tyrosine Kinase
Authors: Stellrecht Christine M. , Phillip Cornel J. , Cervantes-Gomez Fabiola , Gandhi Varsha
Published: March 2023

Abstract
Multiple myeloma (MM) is an invariably fatal plasma cell malignancy, primarily due to the therapeutic resistance which ultimately arises. Much of the resistance results from the expression of various survival factors. Despite this, the ribonucleoside analogue, 8-chloro-adenosine (8-Cl-Ado), is cytotoxic to a number of MM cell lines. Previously, we established that the analogue incorporates into the RNA and inhibits mRNA synthesis. Because 8-Cl-Ado is able to overcome survival signals present in MM cells and inhibits mRNA synthesis, it is likely that the drug induces cytotoxicity by depleting the expression of critical MM survival genes. We investigated this question using gene array analysis, real-time reverse transcription-PCR, and immunoblot analysis on 8-Cl-Ado–treated MM.1S cells and found that the mRNA and protein levels of the receptor tyrosine kinase MET decrease prior to apoptosis. To determine MET's role in 8-Cl-Ado cytotoxicity, we generated MM.1S clones stably expressing a MET ribozyme. None of the clones expressed <25% of the basal levels of MET mRNA, suggesting that a threshold level of MET is necessary for their survival. Additionally, the ribozyme knockdown lines were more sensitive to the cytotoxic actions of 8-Cl-Ado as caspase-3 activation and the induction of poly-ADP-ribose polymerase (PARP) cleavage were more pronounced and evident 12 h earlier than in the parental cells. We further established MET's role in MM cell survival by demonstrating that a retroviral MET RNA interference construct induces PARP cleavage in MM.1S cells. These results show that MET provides a survival mechanism for MM cells. 8-Cl-Ado overcomes MM cell survival by a mechanism that involves the depletion of MET. [Cancer Res 2007;67(20):9913–20]
Data from A Novel Small Molecule Inhibitor of Signal Transducers and Activators of Transcription 3 Reverses Immune Tolerance in Malignant Glioma Patients
Authors: Hussain S. Farzana , Kong Ling-Yuan , Jordan Justin , Conrad Charles , Madden Timothy , Fokt Isabella , Priebe Waldemar , Heimberger Amy B.
Published: March 2023

Abstract
Overcoming the profound immunosuppression in patients with solid cancers has impeded efficacious immunotherapy. Signal transducers and activators of transcription 3 (STAT3) has recently emerged as a potential target for effective immunotherapy, and in this study, we describe a novel small molecule inhibitor of STAT3 that can penetrate the central nervous system (CNS) in mice and in physiologically relevant doses in vitro and reverse tolerance in immune cells isolated from glioblastoma multiforme (GBM) patients. Specifically, it induces the expression of costimulatory molecules on peripheral macrophages and tumor-infiltrating microglia, stimulates the production of the immune-stimulatory cytokines interleukin 2 (IL-2), IL-4, IL-12, and IL-15, and induces proliferation of effector T cells from GBM patients that are refractory to CD3 stimulation. We show that the functional enhancement of immune responses after STAT3 inhibition is accompanied by up-regulation of several key intracellular signaling molecules that critically regulate T-cell and monocyte activation. Specifically, the phosphorylation of Syk (Tyr352) in monocytes and ZAP-70 (Tyr319) in T cells are enhanced by the STAT-3 inhibitor in marked contrast to toll-like receptor and T-cell receptor agonists, respectively. This novel small molecule STAT3 inhibitor has tremendous potential for clinical applications with its penetration into the CNS, easy parental administration, direct tumor cytotoxicity, and potent immune adjuvant responses in immunosuppressed cancer patients. [Cancer Res 2007;67(20):9630–6]
Data from A Novel Gene Expression Profile in Lymphatics Associated with Tumor Growth and Nodal Metastasis
Authors: Clasper Steven , Royston Daniel , Baban Dilair , Cao Yihai , Ewers Stephan , Butz Stefan , Vestweber Dietmar , Jackson David G.
Published: March 2023

Abstract
Invasion of lymphatic vessels is a key step in the metastasis of primary tumors to draining lymph nodes. Although the process is enhanced by tumor lymphangiogenesis, it is unclear whether this is a consequence of increased lymphatic vessel number, altered lymphatic vessel properties, or both. Here we have addressed the question by comparing the RNA profiles of primary lymphatic endothelial cells (LEC) isolated from the vasculature of normal tissue and from highly metastatic T-241/vascular endothelial growth factor (VEGF)-C fibrosarcomas implanted in C57BL/6 mice. Our findings reveal significant differences in expression of some 792 genes (i.e., ≥2-fold up- or down-regulated, P ≤ 0.05) that code for a variety of proteins including components of endothelial junctions, subendothelial matrix, and vessel growth/patterning. The tumor LEC profile, validated by immunohistochemical staining, is distinct from that of normal, inflammatory cytokine, or mitogen-activated LEC, characterized by elevated expression of such functionally significant molecules as the tight junction regulatory protein endothelial specific adhesion molecule (ESAM), the transforming growth factor-β coreceptor Endoglin (CD105), the angiogenesis-associated leptin receptor, and the immunoinhibitory receptor CD200, and reduced expression of subendothelial matrix proteins including collagens, fibrillin, and biglycan. Moreover, we show similar induction of ESAM, Endoglin, and leptin receptor within tumor lymphatics in a series of human head and neck and colorectal carcinomas, and uncover a dramatic correlation between ESAM expression and nodal metastasis that identifies this marker as a possible prognostic indicator. These findings reveal a remarkable degree of phenotypic plasticity in cancer lymphatics and provide new insight into the processes of lymphatic invasion and lymph node metastasis. [Cancer Res 2008;68(18):7293–303]
Data from T-cadherin Supports Angiogenesis and Adiponectin Association with the Vasculature in a Mouse Mammary Tumor Model
Authors: Hebbard Lionel W. , Garlatti Michèle , Young Lawrence J.T. , Cardiff Robert D. , Oshima Robert G. , Ranscht Barbara
Published: March 2023

Abstract
T-cadherin delineates endothelial, myoepithelial, and ductal epithelial cells in the normal mouse mammary gland, and becomes progressively restricted to the vasculature during mammary tumorigenesis. To test the function of T-cadherin in breast cancer, we inactivated the T-cadherin (Cdh13) gene in mice and evaluated tumor development and pathology after crossing the mutation into the mouse mammary tumor virus (MMTV)-polyoma virus middle T (PyV-mT) transgenic model. We report that T-cadherin deficiency limits mammary tumor vascularization and reduces tumor growth. Tumor transplantation experiments confirm the stromal role of T-cadherin in tumorigenesis. In comparison with wild-type MMTV-PyV-mT controls, T-cadherin–deficient tumors are pathologically advanced and metastasize to the lungs. T-cadherin is a suggested binding partner for high molecular weight forms of the circulating, fat-secreted hormone adiponectin. We discern adiponectin in association with the T-cadherin–positive vasculature in the normal and malignant mammary glands and report that this interaction is lost in the T-cadherin null condition. This work establishes a role for T-cadherin in promoting tumor angiogenesis and raises the possibility that vascular T-cadherin-adiponectin association may contribute to the molecular cross-talk between tumor cells and the stromal compartment in breast cancer. [Cancer Res 2008;68(5):1407–16]
Data from The Side Population of Ovarian Cancer Cells Is a Primary Target of IFN-α Antitumor Effects
Authors: Moserle Lidia , Indraccolo Stefano , Ghisi Margherita , Frasson Chiara , Fortunato Elena , Canevari Silvana , Miotti Silvia , Tosello Valeria , Zamarchi Rita , Corradin Alberto , Minuzzo Sonia , Rossi Elisabetta , Basso Giuseppe , Amadori Alberto
Published: March 2023

Abstract
The side population (SP), recently identified in several normal tissues and in a variety of tumors based on its ability to extrude some fluorescent dyes, may comprise cells endowed with stem cell features. In this study, we investigated the presence of SP in epithelial ovarian cancer and found it in 9 of 27 primary tumor samples analyzed, as well as in 4 of 6 cultures from xenotransplants. SP cells from one xenograft bearing a large SP fraction were characterized in detail. SP cells had higher proliferation rates, were much less apoptotic compared with non-SP cells, and generated tumors more rapidly than non-SP cells. We also investigated the effects of IFN-α, a cytokine that has widely been used to treat solid tumors, on epithelial ovarian cancer cells and observed that IFN-α exerted marked antiproliferative and proapoptotic effects on primary cultures containing high numbers of SP cells. In vitro, IFN-α treatment invariably caused a dramatic reduction in SP size in tumor cell lines of different origins; moreover, IFN-α treatment of purified SP cells was associated with a distinctive change in their transcriptional profile. Gene therapy with human IFN-α resulted in regression of established tumors bearing a large SP fraction, which was not observed when tumors bearing low SP levels were treated. These findings could have relevant clinical implications because they imply that tumors bearing large SP numbers, albeit rare, could be sensitive to IFN-α treatment. [Cancer Res 2008;68(14):5658–68]
Data from Knockdown of the Transforming Growth Factor-β Type III Receptor Impairs Motility and Invasion of Metastatic Cancer Cells
Authors: Criswell Tracy L. , Dumont Nancy , Barnett Joey V. , Arteaga Carlos L.
Published: March 2023

Abstract
The transforming growth factor-β (TGF-β) signaling pathway plays dual roles in epithelial cell tumorigenesis. TGF-β is initially growth inhibitory, but as tumorigenesis progresses, TGF-β becomes prometastatic. Although the role of the types I and II TGF-β receptors is fairly well established, the role of the ubiquitously expressed TGF-β type III receptor (TβRIII) in tumorigenesis is less defined. To examine the role of TβRIII in breast cancer cells, we stably expressed short hairpin RNAs specific to TβRIII in MDA-231 human breast cancer cells and mouse mammary carcinoma cells expressing the polyomavirus middle T oncogene (PMTLuc). MDA-231 and PMTLuc cells with down-regulated TβRIII expression (231-kd; PMTLuc-kd) exhibited decreased growth rate, motility, and invasion into Matrigel, as well as an increase in apoptosis, compared with control cells. MDA-231 xenografts established in nude mice metastasized, whereas tumors made by 231-kd cells did not. Nuclear factor-κB (NF-κB) activity, which is known to regulate cell growth and motility, was lower in the MDA-231 and PMTLuc knockdown cells compared with control cells. Transfection of an expression vector encoding constitutively active IKK2 into the 231-kd cells restored the ability of TβRIII-deficient cells to invade Matrigel and decreased their basal level of apoptosis. These data indicate that TβRIII differentially regulates cell growth, motility, and invasion in tumorigenic MDA-231 and PMTLuc cells and that these growth changes occur through the modulation of NF-κB activity. [Cancer Res 2008;68(18):7304–12]
Data from Recruitment and Activation of Natural Killer Cells In vitro by a Human Dendritic Cell Vaccine
Authors: Gustafsson Karin , Ingelsten Madeleine , Bergqvist Linda , Nyström Jenny , Andersson Bengt , Karlsson-Parra Alex
Published: March 2023

Abstract
Recruitment of circulating natural killer (NK) cells into inflamed lymph nodes is known to provide a potent, IFN-γ–dependent boost for Th1-polarized immune responses in mouse models. Such NK cell recruitment into draining lymph nodes is induced by certain s.c. injected adjuvants, including mature vaccine dendritic cells (DC), and is mediated by a CXCR3-dependent pathway. Here, we show that monocyte-derived immature human DCs stimulated with polyinosinic acid:polycytidylic acid, IFN-α, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IFN-γ, α-type 1–polarized DC (αDC1), secrete profuse amounts of the CXCR3 ligand CXCL9/MIG and substantial amounts of CXCL10/IP-10 and CXCL11/I-TAC after withdrawal of maturation stimuli. In sharp contrast, no measurable production of these chemokines was found in DCs after maturation with the current gold standard maturation cocktail for human DC-based cancer vaccines consisting of TNF-α, IL-1β, IL-6, and prostaglandin-E2 (PGE2-DC). PGE2-DCs preferentially produced the Th2 and regulatory T-cell–attracting chemokines CCL17/TARC and CCL22/MDC, whereas only marginal levels of these chemokines were produced by αDC1s. Functional studies in vitro showed that supernatants from mature αDC1s actively recruited CD3−CD56+ NK cells and that adding anti-CXCL9/MIG antibodies to the αDC1 supernatant substantially reduced this recruitment. Finally, αDC1s were able to induce IFN-γ production when cocultured with resting autologous NK cells, but only if concurrent CD40 ligation was provided. These novel findings indicate that injected human αDC1-based vaccines have the potential to recruit and activate NK cells during their arrival to draining lymph nodes and that this feature may be of relevance for efficient priming of Th1 cells and CTLs. [Cancer Res 2008;68(14):5965–71]
Data from A Small Molecule Disruptor of Rb/Raf-1 Interaction Inhibits Cell Proliferation, Angiogenesis, and Growth of Human Tumor Xenografts in Nude Mice
Authors: Kinkade Rebecca , Dasgupta Piyali , Carie Adam , Pernazza Daniele , Carless Melanie , Pillai Smitha , Lawrence Nicholas , Sebti Said M. , Chellappan Srikumar
Published: March 2023

Abstract
Although it is well established that cyclin-dependent kinases phosphorylate and inactivate Rb, the Raf-1 kinase physically interacts with Rb and initiates the phosphorylation cascade early in the cell cycle. We have identified an orally active small molecule, Rb/Raf-1 disruptor 251 (RRD-251), that potently and selectively disrupts the Rb/Raf-1 but not Rb/E2F, Rb/prohibitin, Rb/cyclin E, and Rb/HDAC binding. The selective inhibition of Rb/Raf-1 binding suppressed the ability of Rb to recruit Raf-1 to proliferative promoters and inhibited E2F1-dependent transcriptional activity. RRD-251 inhibited anchorage-dependent and anchorage-independent growth of human cancer cells and knockdown of Rb with short hairpin RNA or forced expression of E2F1 rescued cells from RRD-251–mediated growth arrest. P.o. treatment of mice resulted in significant tumor growth suppression only in tumors with functional Rb, and this was accompanied by inhibition of angiogenesis, inhibition of proliferation, decreased phosphorylated Rb levels, and inhibition of Rb/Raf-1 but not Rb/E2F1 binding in vivo. Thus, selective targeting of Rb/Raf-1 interaction seems to be a promising approach for developing novel chemotherapeutic agents. [Cancer Res 2008;68(10):3810–8]
Data from Profound but Dysfunctional Lymphangiogenesis via Vascular Endothelial Growth Factor Ligands from CD11b+ Macrophages in Advanced Ovarian Cancer
Authors: Jeon Bong-Hyun , Jang Cholsoon , Han Jinah , Kataru Raghu P. , Piao Lianhua , Jung Keehoon , Cha Hye Ji , Schwendener Reto A. , Jang Kyu Yun , Kim Kwan-Sik , Alitalo Kari , Koh Gou Young
Published: March 2023

Abstract
Severe ascites is a hallmark of advanced ovarian cancer (OVCA), yet the underlying mechanism that creates an imbalance between peritoneal vascular leakage and lymphatic drainage is unknown. Here, we identified and characterized peritoneal lymphatic vessels in OVCA mice, a model generated by implantation of human OVCA cells into athymic nude mice. The OVCA mice displayed substantial lymphangiogenesis and lymphatic remodeling, massive infiltration of CD11b+/LYVE-1+ macrophages and disseminated carcinomatosis in the mesentery and diaphragm, and progressive chylous ascites formation. Functional assays indicated that the abnormally abundant lymphatic vessels in the diaphragm were not conductive in peritoneal fluid drainage. Moreover, lipid absorbed from the gut leaked out from the aberrant mesenteric lymphatic vessels. Our results indicate that vascular endothelial growth factor (VEGF)-C, VEGF-D, and VEGF-A from CD11b+ macrophages are responsible for producing OVCA-induced dysfunctional lymphangiogenesis, although other cell types contribute to the increased ascites formation. Accordingly, the combined blockade of VEGF-C/D and VEGF-A signaling with soluble VEGF receptor-3 and VEGF-Trap, respectively, markedly inhibited chylous ascites formation. These findings provide additional therapeutic targets to ameliorate chylous ascites formation in patients with advanced OVCA. [Cancer Res 2008;68(4):1100–9]