siRNA Directed Against Survivin Enhances Pancreatic Cancer Cell Gemcitabine Chemosensitivity
Survivin is known to be overexpressed in various human malignancies, including pancreatic cancer, and to cause resistance to radiation and chemotherapy, so the regulation of this molecule could be a new strategy for treating pancreatic cancer. In our study, a short interfering RNA (siRNA) plasmid expression vector against survivin was constructed and transfected into human pancreatic cancer cell lines of Panc-1 and BxPC3. The expression of survivin mRNA and protein among the stable transfected cells and the untransfected cells was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. Tumor cell growth in vitro was assessed by trypan blue exclusion. The cell cycle distribution and cell apoptosis were measured by flow cytometry. The cytotoxicity assay was measured by the MTT test. Our results showed that survivin siRNA treatment caused a specific and profound decrease of survivin mRNA and protein that was associated with decreased cell growth, spontaneous apoptosis, and a specific G0/G1 arrest. Furthermore, the suppression of survivin can enhance the chemosensitivity of pancreatic cancer cells to gemcitabine significantly. We suggest that the RNAi against survivin gene strategy would be a potential approach to chemosensitization therapy in human pancreatic cancer.
KeywordsPancreatic cancer Chemosensitivity RNA interference Survivin
We thank Jin Huang (Institute of Liver Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, P.R. China) for the technical assistance. This work was supported by the National Natural Science Foundation of China, grant nos. 30471693 and 30600592.
- 7.Satoh K, Kaneko K, Hirota M, Masamune A, Satoh A, Shimosegawa T (2001) Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors. Cancer 92:271–278. doi: 10.1002/1097-0142(20010715)92:2<271::AID-CNCR1319>3.0.CO;2-0PubMedCrossRefGoogle Scholar
- 32.Takizawa BT, Uchio EM, Cohen JJ, Wheeler MA, Weiss RM (2007) Downregulation of survivin is associated with reductions in TNF receptors’ mRNA and protein and alterations in nuclear factor kappa B signaling in urothelial cancer cells. Cancer Invest 25:678–684. doi:10.1080/07357900701600954 PubMedCrossRefGoogle Scholar
- 33.Gazzaniga P, Silvestri I, Gradilone A, Scarpa S, Morrone S, Gandini O, Gianni W, Frati L, Aglianò AM (2007) Gemcitabine-induced apoptosis in 5637 cell line: an in-vitro model for high-risk superficial bladder cancer. Anticancer Drugs 18:179–185. doi:10.1097/CAD.0b013e328010ef47 PubMedCrossRefGoogle Scholar