【摘要】 目的:探究siRNA沉默钙蛋白酶1(Calpain1,CAPN1)对胆囊癌细胞GBC-SD增殖和迁移的影响及其潜在的分子机制。方法:将阴性siRNA、CAPN1-siRNA分别转染至细胞GBC-SD,以未转染的细胞为对照组,转染48 h后,采用荧光倒置显微镜检测转染效率,实时荧光定量PCR(qRT-PCR)检测GBC-SD细胞中CAPN1的表达情况,应用MTT实验检测GBC-SD细胞增殖能力,采用划痕实验检测GBC-SD细胞迁移能力,采用蛋白免疫印迹(Western blot)检测GBC-SD细胞中增殖细胞核抗原(PCNA)和CD44蛋白表达情况。结果:显微镜检测结果显示转染效率达到90%以上;与对照组、siRNA组相比,CAPN1-siRNA组细胞增殖率和迁移能力显著降低,CAPN1、PCNA、CD44蛋白表达显著降低(P <0. 05)。结论:siRNA沉默CAPN1具有一定的抑制胆囊癌细胞增殖和迁移作用,可为胆囊癌的治疗提供新的实验证据。更多还原

【Abstract】 Objective: To explore the effects of siRNA silencing of calpain 1( CAPN1) on the proliferation and migration of gallbladder carcinoma GBC-SD cells and the underlying molecular mechanisms. Methods: CAPN1-siRNA and siRNA were transfected into gallbladder cancer cell GBC-SD,and respectively named as CAPN1-siRNA group and siRNA group. The cells without transfection were served as the control. After 48 hours,a fluorescence microscope was used to detect the transfection efficiency,real-time fluorescence quantitative PCR( qRT-PCR) was used to detect the expression of CAPN1 in GBC-SD cells,MTT and scratch assay were used to detect the proliferation and migration ability of GBC-SD cells,and Western blot was used to detect the expressions of CD44 and PCNA in GBC-SD cells. Results: The results showed that the transfection efficiency reached above 90%; the proliferation and migration rate in CAPN1-siRNA group was significantly lower than that in the control group( P < 0. 05). The expression levels of CAPN1,PCNA and CD44 were significantly lower than those in the control group( P < 0. 05). Conclusion: SiRNA silencing CAPN1 can inhibit the proliferation and migration of gallbladder cancer cells,which may provide new experimental evidence for the treatment of gallbladder cancer.更多还原