CAJ | 학술논문

目的探讨吉西他滨对Beclinl基因沉默的人胰腺癌MiaPaCa-2细胞周期及凋亡的影响.方法构建靶向Beclinl的siRNA,插入表达质粒,转染MiaPaCa-2细胞.采用RT-PCR法和蛋白质印迹法检测细胞Beclinl mRNA及蛋白的表达,应用吉西他滨处理Beclinl基因沉默的MiaPaCa-2细胞,应用流式细胞仪检测细胞周期及细胞凋亡状况.结果成功获得Beclin1基因沉默的MiaPaCa-2细胞,转染后细胞的Beclin1 mRNA表达量从对照组的1.0下降到0.295;S、G2期细胞数减少,而G1期细胞增多;细胞凋亡未受影响.应用吉西他滨处理后,Beclin1基因沉默的MiaPaCa-2细胞的S期细胞数进一步减少,而G1、G2期细胞增多,细胞凋亡被抑制.结论 Beclinl表达沉默使人胰腺癌细胞系MiaPaCa-2细胞周期发生改变,并影响吉西他滨对细胞周期及凋亡的作用.
목적 탐토길서타빈대Beclinl기인침묵적인이선암MiaPaCa-2세포주기급조망적영향.방법 구건파향Beclinl적siRNA,삽입표체질립,전염MiaPaCa-2세포.채용RT-PCR법화단백질인적법검측세포Beclinl mRNA급단백적표체,응용길서타빈처리Beclinl기인침묵적MiaPaCa-2세포,응용류식세포의검측세포주기급세포조망상황.결과 성공획득Beclin1기인침묵적MiaPaCa-2세포,전염후세포적Beclin1 mRNA표체량종대조조적1.0하강도0.295;S、G2기세포수감소,이G1기세포증다;세포조망미수영향.응용길서타빈처리후,Beclin1기인침묵적MiaPaCa-2세포적S기세포수진일보감소,이G1、G2기세포증다,세포조망피억제.결론 Beclinl표체침묵사인이선암세포계MiaPaCa-2세포주기발생개변,병영향길서타빈대세포주기급조망적작용.
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.