Oncology Research 21(2) Abstracts

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Oncology Research, Vol. 21, pp. 67–72, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504013X13747716581417
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Ran GTPase Induces EMT and Enhances Invasion in Non-Small Cell Lung Cancer Cells Through Activation of PI3K-AKT Pathway

Jinfeng Ning,*1 Wei Liu,†1 Jinfeng Zhang,* Yaoguo Lang,* and Shidong Xu*

*Department of Thoracic Surgery, The Affiliated Tumor Hospital of Harbin Medical University, Harbin, PR China
†Department of Breast Medical Oncology, The Affiliated Tumor Hospital of Harbin Medical University, Harbin, PR China

Ras-related nuclear protein (Ran) GTPase is upregulated in non-small cell lung cancer (NSCLC) cells and is required for NSCLC cell survival. However, the effect of Ran on NSCLC cell invasion and epithelial to mesenchymal transition (EMT) remains unclear. This study found that Ran expression was much higher in highly invasive NSCLC cells than in lowly invasive NSCLC cells. Ectopic expression of Ran enhanced invasion and induced EMT in NSCLC cells. Inhibition of the PI3K-AKT pathway by LY294002, but not the MEK-ERK pathway by PD98509, reversed the above effects in these cells induced by Ran overexpression. In conclusion, our findings demonstrate that Ran induces EMT and enhances invasion in NSCLC cells through the activation of PI3K-AKT signaling. Thus, Ran may be a potential target for NSCLC therapeutic intervention.

Key words: Ras-related nuclear protein (Ran); Non-small cell lung cancer (NSCLC); Invasion; Epithelial to mesenchymal transition (EMT); PI3K-AKT signaling

1These authors provided equal contribution to this work.
Address correspondence to Shidong Xu, Department of Thoracic Surgery, The Affiliated Tumor Hospital of Harbin Medical University, Harbin, PR China. Tel and Fax: +86-0451-62938870; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 73–82, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504013X13775486749173
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

RNAi Targeting of CCR2 Gene Expression Induces Apoptosis and Inhibits the Proliferation, Migration, and Invasion of PC-3M Cells

Ji Gao,*1 Anna Wang,†1 Muchun Zhang,*1 Hongyan Li,* Kaichen Wang,* Yuping Han,* Zhixin Wang,* Chaoling Shi,‡ and Weihua Wang*

*Department of Urology, China–Japan Union Hospital, Jilin University, Changchun, Jilin, China
†Department of Hepatology, Jilin Academy of Traditional Chinese Medicine, Changchun, Jilin, China
‡Department of Urology, Fourth Hospital of Jilin University, Changchun, Jilin, China

Prostate cancer (PCa) is the second most lethal malignancy in men. It has been reported that chemokines, produced by cancer cells, have linked with tumor progression and metastatic spread. We have proven that chemokine (C-C) motif ligand 2 (CCL2) is involved in the growth and invasion of PCa. In this study, we studied whether CC chemokine receptor 2 (CCR2), the receptor of CCL2, also contributes to PCa progression. We constructed the recombinant plasmid pGCsi-CCR2 and investigated the effects of pGCsi-CCR2 on proliferation, apoptosis, migration, and invasion of PC-3M cells. RT-PCR and Western blot assay showed that transfection with the plasmid pGCsi-CCR2 successfully inhibited the CCR2 expression. The cell proliferation rate was significantly slow, and the apoptotic rate was increased in PC-3M cells treated with CCR2-siRNA, indicated by MTT cell viability and TUNEL assay, respectively. As expected, CCR2 knockdown also reduced the migration and invasion of PC-3M cells, as illustrated through wound-healing assay and Transwell assay. The possible molecular mechanism was the regulation of several signal pathways involved in survival, apoptosis, migration, and metastasis. Altogether, the present finding suggests that CCR2 expression is crucial for CCL2-induced proliferation and invasion of PC-3M cells, and CCR2 could also be a promising molecular target for prevention of PCa growth and metastasis.

Key words: RNA interference; CC chemokine receptor 2 (CCR2); Prostate cancer cells; Apoptosis; Metastasis

1These authors provided equal contribution to this work.
Address correspondence to Weihua Wang, Department of Urology, China–Japan Union Hospital, Jilin University, Changchun, Jilin 130033, China. Tel/Fax: +86-0431-85281643; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Chaoling Shi, Department of Urology, Fourth Hospital of Jilin University, Changchun, Jilin 130033, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 83–91, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504013X13775486749218
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

MiR-138 Induces Renal Carcinoma Cell Senescence by Targeting EZH2 and Is Downregulated in Human Clear Cell Renal Cell Carcinoma

Jiaqian Liang,*1 Yajing Zhang,†1 Guosong Jiang,* Zhouqiang Liu,* Wei Xiang,* Xuanyu Chen,* Zhaohui Chen,* and Jun Zhao*

*Department of Urology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, PR China
†Department of Nuclear Medicine, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, PR China

MiR-138 has been shown to be downregulated in various cancers, including head and neck squamous cell carcinoma (HNSCC) and clear cell renal carcinoma (ccRCC). In the present study, we aimed to reveal the mechanism of miR-138 induction of senescence in renal carcinoma cells and identify its specific target genes. We used qRT-PCR to analyze miR-138 expression levels in renal carcinoma cell lines and ccRCC samples. The activity of β-galactosidase was measured for functional analysis after miR-138 mimic transfection. To identify the targets of miR-138, we used three types of target prediction software to determine three candidate target genes. Furthermore, a 3′UTR luciferase assay was performed. Western blotting was used to detect the protein expression levels of candidate target genes. Additionally, knockdown of EZH2 by its siRNA was performed. The expression of miR-138 was downregulated in RCC cells lines and in tumor samples compared with their controls. Transfection of miR-138 mimic induced SN-12 cell senescence, decreased the protein expression of EZH2, and increased the protein expression of P16. Furthermore, miR-138 decreased the 3′UTR luciferase activity of EZH2. The knockdown of EZH2 by siRNA induced SN-12 cell senescence, decreased the protein expression level of EZH2, and increased the protein expression of P16. MiR-138 is a tumor-suppressor miRNA in ccRCC that induces SN-12 cell senescence by downregulating EZH2 expression and upregulating P16 expression.

Key words: MiR-138; EZH2; P16; Renal carcinoma; Senescence

1These authors provided equal contribution to this work and should be considered as co-first authors.
Address correspondence to Jun Zhao, Department of Urology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, PR China. Tel: +86-27-85351624, +86-13807165235; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Zhaohui Chen, Department of Urology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, PR China. Tel: +86-27-85351625, +86-18627956868; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 93–102, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504013X13786659070154
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Wnt Antagonist, Secreted Frizzled-Related Protein 4 (sFRP4), Increases Chemotherapeutic Response of Glioma Stem-Like Cells

Sudha Warrier,* Senthil Kumar Balu,* Alan Prem Kumar,†‡§¶ Michael Millward,# and Arunasalam Dharmarajan#1

*Division of Cancer Stem Cells and Cardiovascular Regeneration, Manipal Institute of Regenerative Medicine, Manipal University, Bangalore, India
†Cancer Science Institute of Singapore, National University of Singapore, Singapore
‡Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
§Department of Biological Sciences, University of North Texas, Denton, TX, USA
¶School of Biomedical Sciences, Faculty of Health Sciences, Curtin University, Western Australia, Australia
#School of Anatomy, Physiology and Human Biology, The University of Western Australia, Perth, Western Australia, Australia

Malignant gliomas have a highly tumorigenic subpopulation, termed cancer stem cells (CSCs), that drives tumor formation and proliferation. CSCs possess inherent resistance mechanisms against radiation- and chemotherapy-induced cancer cell death, enabling them to survive and initiate tumor recurrence. We examined the effect of secreted frizzled-related protein 4 (sFRP4), a Wnt signaling antagonist, in chemosensitizing the glioma cell line U138MG and glioma stem cells (GSCs) enriched from U138MG to chemotherapeutics. We found that sFRP4 alone and in combination with either doxorubicin or cisplatin induced apoptosis. Proliferation decreased substantially in GSC-enriched population as measured by MTT and BrdU assays. JC-1 and caspase-3 assays demonstrated that cell death was through the apoptotic pathway. sFRP4 treatment also decreased neurosphere formation and induced neuronal differentiation. Inhibition by sFRP4 was abolished by Wnt3a, indicating that sFRP4 acts through the frizzled receptor. Further indication that sFRP4 acts through the Wnt β-catenin pathway was provided by decrease in the β-catenin protein and decrease in the β-catenin-stimulated gene cyclin D1 upon sFRP4 induction. By real-time PCR, an increase in apoptotic markers Bax and p21, a decrease in pro-proliferative marker CycD1, and a decrease in the GSC marker CD133 were observed. These observations indicate that sFRP4 is able to sensitize glioma cells and stem cells to chemotherapeutics. We thus identified for the first time that sFRP4 could help to destroy cancer stem cells of glioma cell line, which would lead to effective treatment regimen to combat brain tumors.

Key words: Glioma stem cells (GSCs); Secreted frizzled-related protein 4 (sFRP4); Apoptosis; Chemosensitivity; Wnt antagonist

1Current address: School of Biomedical Sciences, Faculty of Health Sciences, Curtin University, Western Australia, Australia.
Address correspondence to Sudha Warrier, Division of Cancer Stem Cells and Cardiovascular Regeneration, Manipal Institute of Regenerative Medicine, Manipal University, Bangalore 560 065, India. Tel: +91-80-28460671, 28460691; Fax: +91-80 28460691; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 103–110, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504013X13808175127324
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

The Insulin-Like Growth Factor-I Receptor Inhibitor Picropodophyllin-Induced Selective Apoptosis of Hepatocellular Carcinoma Cell Through a Caspase-Dependent Mitochondrial Pathway

Changyong E,* Jing Li,† Dan Shao,† Dan Zhang,* Yue Pan,† Li Chen,† and Xuewen Zhang*

*Department of Hepatobiliary and Pancreatic Surgery, China-Japan Union hospital of Jilin University, Changchun, Jilin, China
†Department of pharmacology of Basic Medical Sciences School, Jilin University, Changchun, Jilin, China

The insulin-like growth factor-I receptor (IGF-1R) and its ligands (IGF-I, IGF-II) have been shown to be important promoters of cancer development and are frequently overexpressed in most hepatocellular carcinomas (HCCs). The activation of IGF-1R signaling mediates tumorigenesis, proliferation, and metastasis and thus represents a potential target for innovative treatment strategies for HCC. We investigated the potential inhibitory effect and mechanism of the impact of a novel IGF-1R inhibitor, picropodophyllin (PPP), in HCC lines. It was found that PPP selectively induced cell apoptosis in a time- and dose-dependent manner in HCC cells compared to normal hepatocytes. The inhibitory effects had a positive correlation with the expression of IGF-1R. PPP exerted an apoptotic effect in HCC cells in a caspase-dependent manner through the mitochondrial pathway. The release of cytochrome C from the mitochondrion was coupled with activation of caspase-9 and caspase-3. Treatment of PPP in HepG2 cells resulted in a marked elevation of Bax protein, but decreased levels of phosphorylated Akt and Bcl-2 protein. The ratio of Bax/Bcl-2 was increased in a dose-dependent manner. Our study provides strong evidence that the IGF-1R inhibitor PPP selectively inhibits the growth of human hepatocellular cancer cells by inducing the caspase-dependent mitochondrial pathway cell apoptosis pathway with no observed cytotoxicity on normal cells.

Key words: Hepatocellular carcinoma (HCC); Insulin-like growth factor-I receptor (IGF-1R); Picropodophyllin (PPP); Selective; Apoptosis

Address correspondence to Xuewen Zhang, M.D., Ph.D., Department of Hepatobiliary and Pancreatic Surgery, China–Japan Union Hospital of Jilin University, Changchun 130033, Jilin, China. Tel: +86-0431-84995099; Fax: +86-0431-88796059; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Li Chen, Ph.D., Department of Pharmacology of Basic Medical Sciences School, Changchun 130021, Jilin, China. Tel: +86-0431-84995099; Fax: +86-0431-88796059; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 111–120, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504013X13814233062171
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

FOXC2 Often Overexpressed in Glioblastoma Enhances Proliferation and Invasion in Glioblastoma Cells

Weihua Li,*1 Xin Fu,†1 Rongyao Liu,* Chunming Wu,* Jingyang Bai,* Yousong Xu,* Yongshun Zhao,* and Yinghui Xu*

*Department of Neurosurgery, the First Affiliated Hospital of Dalian Medical University, Dalian, P.R. China
†Dalian Medical University, Dalian, P.R. China

Glioblastoma (GBM) is the most common primary brain tumor and the leading cause of tumor-related death in the central nervous system. To date, the mechanisms of GBM genesis remain elusive. Forkhead box protein C2 (FOXC2) is a transcription factor that has been reported in many cancers, but its function in GBM tumorigenesis is not clearly elucidated. This study found that FOXC2 was overexpressed in GBM cell lines and GBM tissues. The proliferation and invasive potential of GBM cells were significantly increased by ectopic expression of FOXC2 but significantly decreased by RNA interference targeting FOXC2. EGFR expression was modulated by FOXC2 both in mRNA and protein levels. EGFR inhibition by siRNA reversed the FOXC2-induced proliferation and invasion. These findings suggested that FOXC2 expressed in GBM has a function in GBM cell proliferation and invasion and may be partly associated with the EGFR overexpression.

Key words: Forkhead box protein C2 (FOXC2); Glioblastoma (GBM); Proliferation; Invasion; Epidermal growth factor receptor (EGFR)

1These authors provided equal contribution to this work.
Address correspondence to Yinghui Xu, Department of Neurosurgery, the First Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian, 116011, P.R. China. Tel: +86-411-83622844; Fax: +86-411-83622844; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it