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Inhibitory effects of antisense phosphorothioate oligodeoxynucleotides on pancreatic cancer cell Bxpc-3 telomerase activity and cell growth in vitro
Authors:Wang Yun-Feng  Guo Ke-Jian  Huang Bei-Ting  Liu Yong  Tang Xiao-Yun  Zhang Jian-Jun  Xia Qiang
Institution:1. Department of General Surgery, The Center Hospital of Shanghai Yangpu District (Branch Hospital of The Affiliated Xinhua Hospital of Shanghai Jiaotong University),Shanghai 200127, China
2. Department of Biliary-Pancreatic Surgery, The First Affiliated Hospital of China Medical University,Shengyang 110001, Liaoning Province, China
3. Department of Clinical Epidemiology, Staff Room of the First Affiliated Hospital of China Medical University,Shengyang 11001, Liaoning Province, China
4. The First Department of The Center of Experiment and Technology, The China Medical University,Shengyang 11001, Liaoning Province, China
5. Center of Liver Transplantation of The Affiliated Renji Hospital of Shanghai Jiaotong University,Shanghai 200127, China
Abstract:AIM: To investigate the effect of telomerase hTERT gene antisense oligonucleotide (hTERT-ASO) on proliferation and telomerase activity of pancreatic cancer cell line Bxpc-3. METHODS: MTT assay was used to detect the effect of different doses of hTERT-ASO on proliferation of Bxpc-3 cell for different times. To study the anti-tumor activity, the cells were divided into there groups: Control group (pancreatic cancer cell Bxpc-3); antisense oligonucleotide (hTERT-ASO) group; and nosense oligonucleotide group decorated with phosphorothioate. Telomerase activity was detected using TRAP-PCR-ELISA. Cell DNA distribution was examined using flow cytometry assay. Cell apoptosis was observed by transmission electron microscope in each group. RESULTS: After treatment with 6 mmol/L hTERT-ASO, cell proliferation was inhibited in dose- and time-dependent manner. The telomerase activity decreased after treatment with hTERT-ASO for 72 h. Flow cytometry showed the cell number of G0/G1 phase increased from 2.7% to 14.7%, the cell number of S phase decreased from 72.7% to 51.0%, and a sub-Gl stage cell apoptosis peak appeared in front of G1 stage. CONCLUSION: Telomerase antisense oligodeoxy-nucleotide can inhibit the proliferation of pancreatic cancer cell line Bxpc-3 and decrease the telomerase activity and increase cell apoptosis rate in vitro.
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