Study of Inhibiting The Expression of Survivin in Pancreatic Cancer Cells with RNA Interference_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

HE Manxi ¹ , LI Geng ² , YAO Yougui ² , PU Daoshen ² , TIAN Bole ¹ , HU Weiming ¹ , ZHANG Zhaoda ¹

1.Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China;;
2.Department of Hepatobiliary Surgery, People’s Hospital of Ya’an City, Ya’an 625000, China;

Corresponding author：

Keywords：

RNA interference; Pancreatic cancer; Survivin Apoptosis

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Objective To investigate the inhibitory effects of RNA interference (RNAi) expression vector on the expression of survivin in pancreatic cancer cell PANC-1. Methods The protein and mRNA expressions of survivin were examined with immunofluorescence and RT-PCR. The survivin gene was cloned into the T-vector and sequenced. The RNAi expression vectors targeting survivin, named si-svv-1 and si-svv-2 respectively according to whether they harbored a mutation or no mutation, were constructed and transfected into PANC-1 cells with liposome. The expression of survivin mRNA was detected with RT-PCR. Apoptosis of PANC-1 cells was analyzed with DNA ladder and FACS. Results There was a high degree expression of survivin in PANC-1 cells. The expression of survivin was not inhibited by RNAi expression vectors si-svv-1, but inhibited about (72.43±8.04)% by si-svv-2 and the apoptosis rate of PANC-1 cells increased to （12.36±1.44）% after 72 h. Conclusion The RNAi expression vector can effectively inhibit the expression of survivin in pancreatic cancer cell PANC-1 cells and induce the apoptosis in PANC-1 cells.

Citation： HE Manxi,LI Geng,YAO Yougui,PU Daoshen,TIAN Bole,HU Weiming,ZHANG Zhaoda. Study of Inhibiting The Expression of Survivin in Pancreatic Cancer Cells with RNA Interference. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2008, 15(11): 829-834. doi: Copy

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18.	Lu B, Mu Y, Cao C, et al. Survivin as a therapeutic target for radiation sensitization in lung cancer [J]. Cancer Res, 2004; 64(8)∶2840.
19.	Fulda S, Debatin KM. Sensitization for tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by the chemopreventive agent resveratrol [J]. Cancer Res, 2004; 64(1)∶337.
20.	唐朝晖, 邹声泉, 余彦, 等．不同剂量的X射线辐照对胰腺癌细胞Survivin表达的影响 [J]. 临床外科杂志， 2007； 15（5）∶328.
21.	Williams NS, Gaynor RB, Scoggin S, et al. Identification and validation of genes involved in the pathogenesis of colorectal cancer using cDNA microarrays and RNA interference [J]. Clin Cancer Res, 2003; 9(3)∶931.
22.	Fire A, Xu S, Montgomery MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans[J]. Nature, 1998; 391(6669)∶806.
23.	Miyagishi M, Taira K. Development and application of siRNA expression vector [J]. Nucleic Acids Res Suppl, 2002; 2(2)∶113.

1. Rubin P， Williams JP 主编．孙燕, 汤钊猷主译. 临床肿瘤学 [M]. 第8版．北京：人民卫生出版社， 2006∶768.
2. 陆星华．胰腺癌研究现状与展望 [J]. 美中医学， 2005； 2（3）∶73.
3. MacKenzie MJ. Molecular therapy in pancreatic adenocarcinoma [J]. Lancet Oncol, 2004; 5(9)∶541.
4. Lieberman J, Song E, Lee SK, et al. Interfering with disease: opportunities and roadblocks to harnessing RNA interference [J]. Trends Mol Med, 2003; 9(9)∶397.
5. Olie RA, Simes-Wüst AP, Baumann B, et al. A novel antisense oligonucleotide targeting survivin expression induces apoptosis and sensitizes lung cancer cells to chemotherapy [J]. Cancer Res, 2000; 60(11)∶2805.
6. 何满西，孟令祥，唐勇，等． RNAi抑制胰腺癌细胞survivin表达并诱导细胞凋亡的实验研究 [J]. 四川大学学报（医学版）， 2006； 37（4）∶520.
7. Tamm I, Wang Y, Sausville E, et al. IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs [J]. Cancer Res, 1998; 58(23)∶5315.
8. Islam A, Kageyama H, Takada N, et al. High expression of Survivin, mapped to 17q25, is significantly associated with poor prognostic factors and promotes cell survival in human neuroblastoma [J]. Oncogene, 2000; 19(5)∶617.
9. 李哲夫，陈孝平，张万广. 肝细胞癌合并门静脉癌栓组织中PDGF和survivin表达的意义 [J]. 中国普外基础与临床杂志, 2007； 14（1）∶46.
10. 谢贻祥，孟刚. survivin在乳腺癌中的表达及其意义 [J]. 中国普外基础与临床杂志, 2007； 14（3）∶263.
11. Wang Z, Xie Y, Wang H. Changes in survivin messenger RNA level during chemotherapy treatment in ovarian cancer cells[J].Cancer Biol Ther, 2005; 4(7)∶716.
12. Van Valen F, Fulda S, Schfer KL, et al. Selective and nonselective toxicity of TRAIL/Apo2L combined with chemotherapy in human bone tumour cells vs. normal human cells [J]. Int J Cancer, 2003; 107(6)∶929.
13. 陈曦海，张岂凡，马荣. 反义survivin诱导胃癌SGC7901细胞凋亡及逆转耐药的研究 [J].中国普外基础与临床杂志, 2007； 14（1）∶63.
14. Satoh K, Kaneko K, Hirota M, et al. Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors [J]. Cancer, 2001; 92(2)∶271.
15. Jinfeng M, Kimura W, Sakurai F,et al. Histopathological study of intraductal papillary mucinous tumor of the pancreas: special reference to the roles of Survivin and p53 in tumorigenesis of IPMT [J]. Int J Gastrointest Cancer, 2002; 32(2-3)∶73.
16. 王亚利，宋天保，王西京，等． Survivin mRNA反义寡核苷酸诱导胰腺癌细胞凋亡 [J]. 世界华人消化杂志， 2004； 12(8)∶1872.
17. 戴德坚, 陆才德, 赖日勇, 等．靶向抑制survivin对胰腺癌细胞增殖和凋亡效应的影响 [J]. 中华医学杂志, 2004； 84（22）∶1894.
18. Lu B, Mu Y, Cao C, et al. Survivin as a therapeutic target for radiation sensitization in lung cancer [J]. Cancer Res, 2004; 64(8)∶2840.
19. Fulda S, Debatin KM. Sensitization for tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by the chemopreventive agent resveratrol [J]. Cancer Res, 2004; 64(1)∶337.
20. 唐朝晖, 邹声泉, 余彦, 等．不同剂量的X射线辐照对胰腺癌细胞Survivin表达的影响 [J]. 临床外科杂志， 2007； 15（5）∶328.
21. Williams NS, Gaynor RB, Scoggin S, et al. Identification and validation of genes involved in the pathogenesis of colorectal cancer using cDNA microarrays and RNA interference [J]. Clin Cancer Res, 2003; 9(3)∶931.
22. Fire A, Xu S, Montgomery MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans[J]. Nature, 1998; 391(6669)∶806.
23. Miyagishi M, Taira K. Development and application of siRNA expression vector [J]. Nucleic Acids Res Suppl, 2002; 2(2)∶113.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Study of Inhibiting The Expression of Survivin in Pancreatic Cancer Cells with RNA Interference

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