Vav3对Lewis细胞增殖与凋亡影响观察

Effect of siRNA-mediated Vav3 silence on proliferation and apoptosis of Lewis lung carcinoma cells

目的:探讨Vav3调节Lewis肺癌细胞增殖及凋亡的作用及机制。方法:小分子干扰RNA(small interfering RNA,siRNA)下调Lewis肺癌细胞中Vav3蛋白的表达;蛋白质印迹法检测转染siRNA-Vav3后Vav3及JNK蛋白表达的变化;MTT方法检测细胞存活率,流式细胞仪检测细胞凋亡。结果:转染siRNA后,干扰组Vav3表达为0.154±0.024,较Lewis肺癌细胞组的0.417±0.073和对照组的0.383±0.049明显下调,F=22.052,P=0.002。下调Vav3表达后,干扰组JNK蛋白表达水平为0.619±0.008,与肺癌细胞组的1.251±0.117和对照组的1.192±0.092相比较明显下调,F=54.770,P<0.001。与肺癌细胞组细胞生存率(100.00±0.00)%和对照组(95.33±3.14)%相比,干扰组(75.87±2.30)%明显下降,P值分别为0.000 2和0.000 3。与肺癌细胞组细胞凋亡率(3.55±0.03)%和对照组(4.18±0.17)%相比,干扰组(12.16±0.02)%明显上升,P值分别为0.000 1和0.000 1。结论:下调Vav3表达,抑制Lewis肺癌细胞的增殖,并且促进肺癌细胞凋亡;Vav3发挥作用可能与JNK信号激活有关。

OBJECTIVE：To investigate the mechanism and effects of Vav3 on proliferation and apoptosis of Lewis lung carcinoma cells. METHODS：The small interfering RNA （siRNA） of Vav3 （siRNA-Vav3） was transfected into Lewis lung carcinoma ceils to inhibit the basal Vav3 level. The protein levels of Vav3 and JNK were detected by Western blot af- ter knockdown of endogenous Vav3. The cellular proliferation and apoptosis were analyzed by methylthiazolete trazolium （MTT） and flow cytometry（FCM）, respectively. RESULTS： The Vav3 protein expression level of the experimental group, LLC group and control group was 0. 154 ± 0. 024,0. 417 ± 0. 073 and 0. 383 ± 0. 049, respectively after transient transfection of siRNA-Vav3. Analysis of variance showed that there was a significant difference among three groups （F= 22. 052,P=0. 002）. JNK protein expression level of the experimental group, LLC group and control group was 0. 619 ± 0. 008,1.251± 0.117 and 0. 383 ± 0. 049, respectively after transient transfection of siRNA-Vav3. There was a significant difference among three groups（F= 54. 770, P〈0. 001）. The survival rate of cells in expriment group（75.87± 2.30） was decreased than that in LLC group （100.00±0.00）% and in negative control group （95.33 ± 3.14）% （P= 0. 000 2, P=0. 000 3）. In addition,the apoptosis rate of cells in expriment group （12.16±0.02）% was decreased compared to that in LLC group（3.55±0.03） % and in control group （4.18±0.17） %（P=0. 000 1 ,P=0. 000 1）. CONCLUSION：It indica- ted that down-regulated expression of Vav3 protein can inhibit LLC cell proliferation and promote apoptosis of LLC cell, which may be related to the activation of JNK.