WISP-2siRNA对人脑胶质瘤U251细胞增殖、凋亡的影响

Effects of WISP-2 siRNA on proliferation and apoptosis of human glioma U251 cells

摘要:

目的 探讨siRNA沉默WISP-2基因表达对人脑胶质瘤细胞U251增殖、凋亡的影响。方法 脂质体介导siRNA转染人脑胶质瘤细胞U251后，检测U251细胞的增殖、凋亡的变化，并用Western blot法检测WISP-2、Bcl-2、Bax蛋白的表达。结果 WISP-2 siRNA 能有效抑制U251细胞株的生长，诱导凋亡，siRNA干扰组WISP-2蛋白表达水平为（18.67±1.40）%，明显低于空白对照组（70.18±1.82）%和阴性对照组（69.41±1.77）%，且差异有统计学意义（P<0.01）； siRNA干扰组 Bcl-2、Bax 蛋白表达水平为（29.67±1.47）%、（31.62±1.32）%，明显低于空白对照组和阴性对照组，且差异有统计学意义（P<0.01）。结论 WISP-2 siRNA抑制WISP-2 mRNA和蛋白表达后，能有效抑制胶质瘤细胞的增殖，增加U251细胞凋亡，可能通过下调Bcl-2/Bax蛋白表达的比值来诱导细胞凋亡。

Abstract:

Objective To investigate the effects of siRNA-mediated WISP-2 gene silencing on the proliferation and apoptosis of human glioma cell line U251. Methods Liposome-mediated transfection with WISP-2 siRNA was performed in human glioma U251 cells. The effects of WISP-2 suppression on the proliferation and apoptosis of U251 cells were evaluated by MTT assay and flow cytometry. The protein expression of WISP-2, Bcl-2, and Bax after the gene silencing of WISP-2 in U251 cells was measured by Western blot. Results The MTT assay showed that the cell growth rate was significantly lower in siRNA interference group than in blank control group and negative control group (P<0.05). The flow cytometry showed that the mean apoptotic rate of siRNA interference group was 16.59±1.40%, significantly lower than those of blank control group (3.13±0.34%) and negative control group (3.42±0.48%) (P<0.01). WISP-2 siRNA significantly inhibited the growth and induced the apoptosis of U251 cells. WISP-2 protein expression was significantly lower in siRNA interference group (18.67±1.40%) than in blank control group (70.18±1.82%) and negative control group (69.41±1.77%) (P<0.01). The protein expression of Bcl-2 and Bax was significantly lower in siRNA interference group (29.67±1.47% and 31.62±1.32%) than in blank control group and negative control group (P<0.01). Conclusions WISP-2 siRNA, which inhibits the mRNA and protein expression of WISP-2, can effectively suppress the proliferation and promote the apoptosis of U251 cells. And the cell apoptosis may be induced by down-regulating the Bcl-2/Bax ratio.