Effects of gemcitabine on the ceil cycle and apoptosis of pancreatic cancer cells MiaPaCa-2 with Beclinl gene silencing
Hongyu Li, Xiaozhong Guo
Abstract
Hongyu Li, Xiaozhong Guo
Abstract
Objective To investigate the effect of gemcitabine on the cell cycle and apoptosis of the human pancreatic cancer cell line MiaPaCa-2 with Beclinl gene silencing.Methods siRNA targeting at Beclinl gene was constructed,then it was inserted into an expression vector and transfected into MiaPaCa-2 cells.The Beclinl mRNA and protein expression were detected by RT-PCR and Western blot.Gemcitabine was used to treat MiaPaCa-2 with Beclinl gene silencing,then the cell cycle and apoptosis were detected by flow cytometry.Results The MiaPaCa-2 cells with Beclinl gene silencing were successfully constructed,and the expression of Beclin1 mRNA was decreased from 1.0 in control group to 0.295,and number of cells in S and G2 phase was decreased,but number of cells in G1 phase was increased,and there was no change in apoptosis.After gemcitabine treatment,number of cells in S phase was further decreased,but number of cells in G1,G2 phase was increased,and apoptosis was inhibited.Conclusions Beclinl gene silencing can change the cell cycle of pancreatic cancer cells MiaPaCa-2,and influence the effects of gemcitabine on cell cycle and apoptosis. Key words: Pancreatic neoplasms; RNA interference; Gene silencing; Beclinl; Gemcitabine ; Cell cycle; Apoptosis
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Objective To investigate the effect of gemcitabine on the cell cycle and apoptosis of the human pancreatic cancer cell line MiaPaCa-2 with Beclinl gene silencing.Methods siRNA targeting at Beclinl gene was constructed,then it was inserted into an expression vector and transfected into MiaPaCa-2 cells.The Beclinl mRNA and protein expression were detected by RT-PCR and Western blot.Gemcitabine was used to treat MiaPaCa-2 with Beclinl gene silencing,then the cell cycle and apoptosis were detected by flow cytometry.Results The MiaPaCa-2 cells with Beclinl gene silencing were successfully constructed,and the expression of Beclin1 mRNA was decreased from 1.0 in control group to 0.295,and number of cells in S and G2 phase was decreased,but number of cells in G1 phase was increased,and there was no change in apoptosis.After gemcitabine treatment,number of cells in S phase was further decreased,but number of cells in G1,G2 phase was increased,and apoptosis was inhibited.Conclusions Beclinl gene silencing can change the cell cycle of pancreatic cancer cells MiaPaCa-2,and influence the effects of gemcitabine on cell cycle and apoptosis. Key words: Pancreatic neoplasms; RNA interference; Gene silencing; Beclinl; Gemcitabine ; Cell cycle; Apoptosis
Key concepts: Gene silencing, Cell cycle, Gemcitabine, Apoptosis, Transfection, Pancreatic cancer, RNA interference, Cancer research