Oncology Research 21(6) Abstracts

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Oncology Research, Vol. 21, pp. 287–293, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504014X13946388748956
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Regulation of MET Kinase Inhibitor Resistance by Copy Number of MET in Gastric Carcinoma Cells

Yohei Funakoshi,* Toru Mukohara,*‡ Roudy Chiminch Ekyalongo,* Hideo Tomioka,* Yu Kataoka,* Yohei Shimono,*† Naoko Chayahara,* Masanori Toyoda,* Naomi Kiyota,* Yutaka Fujiwara,* and Hironobu Minami*‡

*Division of Medical Oncology/Hematology, Department of Medicine, Kobe University Graduate School of Medicine, Kobe, Japan
†Division of Molecular and Cellular Biology, Department of Biochemistry and Molecular Biology, Kobe University Graduate School of Medicine, Kobe, Japan
‡Cancer Center, Kobe University Hospital, Kobe, Japan

We previously established acquired resistant models for MET-tyrosine kinase inhibitors (TKIs) by continuously exposing the MET-amplified gastric cancer cell line MKN45 to MET-TKIs, PHA665752 (MKN45-PR), or GSK1363089 (MKN45-GR). We found resistant mechanisms caused by increased copy number of MET in both lines and Y1230H mutation in MKN45-PR. We also found that excessive MET signaling caused by these MET alterations resulted in intra-S-phase arrest in the absence of MET-TKIs, so that cells grew faster in the presence of MET-TKIs, a phenomenon referred to as “addiction.” In this study, to investigate reversibility of the acquired resistance and “addiction” to MET-TKIs and their causative MET alterations, we sequentially cultured MKN45-PR and MKN45-GR in decreasing concentrations of MET-TKIs until they were able to grow in a drug-free condition. These “revertant” cell lines (designated MKN45-PR-RE and MKN45-GR-RE) were comparatively analyzed. Growth assay showed that both MKN45-PR-RE and MKN45-GR-RE partially lost the property of “addiction” to MET-TKIs. MKN45-GR-RE lost the property of resistance to GSK1363089, but MKN45-PR-RE retained resistance to PHA665752. Copy numbers and expression and phosphorylation of MET protein reduced in both MKN45-PR-RE and MKN45-GR-RE compared with MKN45-PR and MKN45-GR, respectively, but Y1230H mutation and biochemical resistance to PHA665752 remained in MKN45-PR-RE. The “addiction” to MET-TKIs appeared attributable to increased copy number, and the property and the MET alteration were reversible. The Y1230H mutation appeared enough in itself to keep cells resistant to MET-TKIs and was irreversible.

Key words: Acquired resistance; Addiction; Reversibility; Gastric cancer; MET inhibitor

Address correspondence to Toru Mukohara, M.D., D.Med.Sci.,7-5-2 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan. Tel: +81-78-382-5820; Fax: +81-78-382-5821; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 295–305, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504014X13946388749036
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Activation of uPAR Is Required for Cigarette Smoke Extract-Induced Epithelial–Mesenchymal Transition in Lung Epithelial Cells

Qin Wang,1 Hongchao Wang,1 Yi Zhang, Yuke Zhang, and Wei Xiao

Department of Respiratory Medicine, Qilu Hospital, Shandong University, Jinan, P.R. China

Cigarette smoke is a major risk factor for lung cancer, which may contribute to lung cancer invasion and metastasis. However, the mechanism remains unclear. Epithelial–mesenchymal transition (EMT) is a critical phenotypic alteration of cells that triggers invasion and metastasis. The urokinase-type plasminogen activator receptor (uPAR) is originally thought to assist the directional invasion of migrating cells, and increasing evidences show that overexpression of uPAR in cancer cells promotes EMT. Therefore, we intend to study the role of uPAR in cigarette smoke extract (CSE)-induced EMT in lung epithelial cells. In this study, we showed that lung epithelial cells cultured after CSE treatment demonstrated changes consistent with EMT. E-cadherin was decreased, while vimentin, N-cadherin, and α-SMA expression was increased in both A549 and BEAS-2B cells. Cells acquired a mesenchymal-like morphology and increased cell motility and invasion. In addition, CSE-induced EMT was accompanied by increased expression of uPAR and activation of AKT downstream of uPAR. CSE-induced EMT and activation of AKT were blocked by uPAR gene silencing. Antagonizing PI3K also inhibits development of CSE-induced EMT. We conclude that CSE can induce EMT, and the activity of uPAR-dependent signal pathway in EMT is recapitulated in lung epithelial cells in vitro.

Key words: Cigarette smoke extract (CSE); Epithelial–mesenchymal transition (EMT); Urokinase-type plasminogen activator receptor (uPAR); Lung cancer

1These authors provided equal contribution to this work.
Address correspondence to Wei Xiao, Department of Respiratory Medicine, Qilu Hospital, Shandong University, 107# Wenhua Xi Road, Jinan 250012, P. R. China. Fax: +86 531 82169506; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 307–316, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504014X13983417587320
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Epigenetically Modified Pancreatic Carcinoma PANC-1 Cells Can Act as Cancer Vaccine to Enhance Antitumor Immune Response in Mice

Yifeng Tao,*1 Feng Lin,†1 Tao Li,* Junjie Xie,* Chuan Shen,* and Zhecheng Zhu*

*Organ Transplantation Center, Department of General Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China
†Department of General Surgery, Taizhou First People’s Hospital, Taizhou, Zhejiang, China

Pancreatic cancer is characterized as a type of gastrointestinal tumor with a poor prognosis and high degree of malignancy. CIITA gene was found highly methylated in pancreatic carcinoma cell line PANC-1 and responsible for the low expression of major histocompatability complex II (MHC-II) that may lead to immune evasion. Here, we prepared pancreatic cancer vaccine with PANC-1 cells via epigenetic modification to enhance the MHC-II expression. Then the vaccine was injected into C57BL/6J mice and the effect was examined. Our study found that the vaccine could promote the proliferation of antigen-specific T cells, enhance the killing activity of cytotoxic T lymphocytes (CTLs), promote Th1-type cell-mediated secretion of cytokines IFN-γ and IL-2 while inhibiting Th2-type cell-mediated secretion of IL-4, and inhibit the secretion of TGF-β. Generally, the epigenetically modified vaccine could enhance the body’s antitumor immune response, providing feasibility research on cancer vaccine for therapy of pancreatic cancer.

Key words: Pancreatic cancer; Vaccine; Epigenetic modification; CIITA; Major histocompatability complex II (MHC-II); CD8+ cytotoxic T lymphocyte (CD8+ CTL)

1These authors provided equal contribution to this work.
Address correspondence to Zhecheng Zhu, M.D., Associate Professor, Organ Transplantation Center, Department of General Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, No. 197, Ruijiner Road, Shanghai 200025, China. Tel: +86-21-34187696; Fax: +86-21-34187696; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 317–323, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504014X13983417587401
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

A Phase II Trial of Endostar Combined With Gemcitabine and Cisplatin Chemotherapy in Patients With Metastatic Nasopharyngeal Carcinoma (NCT01612286)


Ting Jin,1 Bin Li,1 and Xiao-Zhong Chen

Department of Radiation Oncology, Zhejiang Cancer Hospital, Hangzhou, Zhejiang, People’s Republic of China

Despite the efficacy of gemcitabine-cisplatin (GC) regimens, the outcome of patients with metastatic nasopharyngeal carcinoma (M NPC) is poor. We conducted a phase II trial to determine the safety and efficacy of Endostar, an endogenous inhibitor of angiogenesis, in combination with GC chemotherapy. A total of 30 patients with M NPC were enrolled. The treatment regimen was a combination of gemcitabine (1,000 mg/m2) on days 1 and 8, cisplatin (80 mg/m2) on day 1, and Endostar (15 mg/day) from day 1 to day 14 of a 21-day cycle for a maximum of four cycles. The primary endpoint was progression-free survival (PFS). The median follow-up was 13.1 months (range: 2.9–20.7 months). A total of 28 patients were evaluated. The median PFS was 19.4 months (95% CI, 13.6–25.1 months). The 1-year PFS rate was 69.8%. The confirmed objective response rate was 85.7% (95% CI, 66.4–95.3%), including complete response in 14 patients (50%). The 1-year overall survival rate was 90.2%. The most common grade 3/4 adverse events were neutropenia (46.4%) and thrombocytopenia (14.3%). Our results suggest that a combination of Endostar with GC chemotherapy can lead to effective tumor regression, control disease progression, and improve prognosis in M NPC. Therefore, a combined Endostar and GC regimen should be considered as a potential treatment for patients with M NPC.

Key words: Nasopharyngeal neoplasms; Endostar; Cisplatin; Gemcitabine; Metastasis

1These authors provided equal contribution to this work.
Address correspondence to Dr. Xioa-Zhong Chen, Department of Radiation Oncology, Zhejiang Cancer Hospital, 38 Guang Ji Road, Hangzhou 310022, China. Tel/Fax: +86-571-88122098; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 325–331, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504014X14024160459087
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Activation of p53 by Sodium Selenite Switched Human Leukemia NB4 Cells From Autophagy to Apoptosis

Zhushi Li, Kejian Shi, Liying Guan, Qian Jiang, Yang Yang, and Caimin Xu

State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences and School of Basic Medicine, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China

It was revealed by our previous research that sodium selenite repressed autophagy accompanied by the induction of apoptosis in human leukemia NB4 cells. The inhibition of autophagy exerted a facilitative effect on apoptosis. In the present study, we further explored the mechanisms underlying the switch from autophagy to apoptosis and elucidated p53 played a key role. Selenite induced phosphorylation of p53 at the vital site Ser15 via p38MAPK and ERK. Subsequently p53 dissociated with its inhibitory protein mouse double minute 2 (MDM2). Meanwhile, the nucleolar protein B23 transferred from the nucleolus to the nucleoplasm and associated with MDM2, probably stabilizing p53. The active p53 participated in the decrease of autophagic protein Beclin-1 and LC-3, as well as activation of apoptosis-related caspases. Furthermore, in p53 mutant U937 leukemia cells, selenite could not elicit such a switch from autophagy to apoptosis, laying emphasis on the crucial role p53 played in this process.

Key words: Apoptosis; Autophagy; p53; MDM2; Sodium selenite

Address correspondence to Caimin Xu, State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences and School of Basic Medicine, Peking Union Medical College and Chinese Academy of Medical Sciences, 5 Dongdan Santiao, Beijing 100005, China. Tel/Fax: +86-10-69156445; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 333–343, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504014X14024160459249
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Anoikis Induction and Inhibition of Peritoneal Metastasis of Pancreatic Cancer Cells by a Nuclear Factor-κB Inhibitor, (–)-DHMEQ

Masanori Sato,* Kazuaki Nakanishi,* Sanae Haga,† Masato Fujiyoshi,* Motoi Baba,* Kazuhiro Mino,* Yimin,‡ Haruki Niwa,§ Hideki Yokoo,* Kazuo Umezawa,¶ Yoshihiro Ohmiya,# Toshiya Kamiyama,* Satoru Todo,** Akinobu Taketomi,* and Michitaka Ozaki†

*Department of Gastroenterological Surgery I, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido, Japan
†Laboratory of Molecular and Functional Bio-Imaging (LMFBI), Hokkaido University Graduate School of Health Sciences, Sapporo, Hokkaido, Japan
‡Department of Advanced Medicine, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido, Japan
§Department of Engineering Science, The University of Electro-Communications Graduate School of Informatics and Engineering, Chofu, Tokyo, Japan
¶Department of Molecular Target Medicine Screening, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan
#Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan
**St. Mary’s Hospital, Kurume, Fukuoka, Japan

The transcription factor nuclear factor-κB (NF-κB) plays a crucial role in pancreatic cancer (PC) progression. NF-κB is also involved in resistance to anoikis, a special type of apoptosis induced when cells are detached from the extracellular matrix or other cells. Anoikis resistance is related to the metastatic abilities of tumor cells; however, little is known about anoikis induction as it relates to inhibition of PC metastasis by NF-κB inhibitors. Here we used a specific NF-κB inhibitor, (−)-dehydroxymethylepoxyquinomicin (DHMEQ), to investigate anoikis induction and peritoneal metastasis suppression following NF-κB inhibition. We transduced Gluc, a secretory form of luciferase, into a PC cell line, AsPC-1 (AsPC-1-Gluc), for our in vivo experiments. (−)-DHMEQ induced anoikis in AsPC-1-Gluc cells as measured by cell survival assays and flow cytometry. The DNA-binding activity of p65 was enhanced immediately after cell detachment from culture dishes in ELISA assays. Some antiapoptotic proteins such as cellular inhibitor of apoptotic protein-1 were consequently upregulated on Western blots. (−)-DHMEQ prevented this increase in p65 activity and the subsequent expressions of antiapoptotic molecules. In a murine xenograft model, anoikis-resistant PC cell lines tended to metastasize to the peritoneum more than anoikis-sensitive cells, suggesting a correlation between anoikis sensitivity and peritoneal metastasis. (−)-DHMEQ successfully inhibited peritoneal metastasis of AsPC-1-Gluc cells. We monitored metastasis inhibition by ex vivo chemiluminescent detection of Gluc secreted from tumor cells into murine plasma and by in vivo imaging. Our results suggest that (−)-DHMEQ inhibited peritoneal dissemination by preventing anoikis resistance of PC cells.

Key words: Anoikis; Dehydroxymethylepoxyquinomicin (DHMEQ); Nuclear factor-κB (NF-κB); Pancreatic cancer (PC); Peritoneal metastasis

Address correspondence to Michitaka Ozaki, Laboratory of Molecular and Functional Bio-Imaging (LMFBI), Hokkaido University Graduate School of Health Sciences, Kita-ku, Kita 12, Nishi 5, Sapporo, Hokkaido 060-0812, Japan. Tel: +81-11-706-3337; Fax: +81-11-706-3337; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 345–352, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504014X14024160459285
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Targeting Smad2 and Smad3 by miR-136 Suppresses Metastasis-Associated Traits of Lung Adenocarcinoma Cells

Yi Yang,*† Lei Liu,*‡ Junchao Cai,*‡ Jueheng Wu,*‡ Hongyu Guan,§ Xun Zhu,*‡ Jie Yuan,*¶ Shengping Chen,* and Mengfeng Li*‡

*Key Laboratory of Tropical Disease Control (Sun Yat-Sen University), Ministry of Education, Guangzhou, Guangdong, China
†Department of Pharmacology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, China
‡Department of Microbiology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, China
§Department of Endocrinology and Diabetes Center, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong, China
¶Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong, China

TGF-β/Smad signaling induces epithelial–mesenchymal transition (EMT) and tumor metastasis. As essential mediators in this pathway, Smad2 and Smad3 have been extensively studied and found to promote EMT and the subsequent mobility as well as invasiveness of lung cancer cells. In the present study, we determined that miR-136 directly targeted Smad2 and Smad3, leading to reduced migration and invasiveness of lung adenocarcinoma (ADC) cell lines, accompanied by increased epithelial markers as well as decreased mesenchymal markers. Moreover, ectopic expression of either Smad2 or Smad3 partially restored the malignant phenotype of ADC cells overexpressing miR-136. Taken together, our data demonstrate that miR-136 may play a tumor-suppressive role by repressing EMT and prometastatic traits via targeting Smad2 and Smad3. The potent antimetastasis property of miR-136 and its multitarget mechanism provide new insights in developing novel therapeutic approaches.

Key words: Lung cancer; miR-136; Smad2; Smad3

Address correspondence to Mengfeng Li, Sun Yat-sen University Zhongshan School of Medicine, 74 Zhongshan Road II, Guangzhou, Guangdong 510080, China. Tel: +86-20-87332748; Fax: +86-20-87331209; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Oncology Research, Vol. 21, pp. 353–363, 2014
0965-0407/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096504014X14024160459122
E-ISSN 1555-3906
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

miR-133a Functions as a Tumor Suppressor and Directly Targets FSCN1 in Pancreatic Cancer

Yong Qin,*†1 Xiaoyan Dang,‡1 Wei Li,* and Qingyong Ma*

*Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University College of Medicine, Xi’an, Shaanxi, China
†Department of General Surgery, The Affiliated Xi’an Central Hospital of Xi’an Jiaotong University College of Medicine, Xi’an, Shaanxi, China
‡Department of Emergency, The Second Affiliated Hospital of Xi’an Jiaotong University College of Medicine, Xi’an, Shaanxi, China

MicroRNA-133a has been proven downregulated in many human malignancies and correlated with tumor progression. However, the roles of miR-133a and its related molecular mechanisms in pancreatic cancer are still not clear. Here we found that miR-133a expression was significantly downregulated in pancreatic cancer tissue samples and cell lines by using quantitative real-time RT-PCR. Decreased miR-133a expression was significantly correlated with aggressive clinicopathological features and poor survival. In addition, miR-133a was identified to be a tumor suppressor, as transfection of miR-133a mimics in PANC-1 cells was able to reduce cell proliferation, invasion, and migration and promote cell apoptosis in vitro and suppress tumorigenicity in vivo. Further, we observed an obvious inverse correlation between FSCN1 and miR-133a levels in tumor samples, and FSCN1 was confirmed as a direct target of miR-133a by using Luciferase Reporter Assay. These findings suggest an important role of miR-133a in the molecular etiology of cancer and implicate its potential application in gene therapy of pancreatic cancer.

Key words: MicroRNA-133a; Pancreatic cancer; FSCN1; Proliferation; Apoptosis; Invasion

1These authors provided equal contribution to this work.
Address correspondence to Qingyong Ma, M.D., Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University College of Medicine, Xi’an 710061, Shaanxi, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it