CXCR4基因沉默对膀胱癌T24细胞增殖和凋亡及迁移能力的影响

The Effect of Inhibition of CXCR4 Gene by siRNA on the Proliferation,Apoptosis and Migration of T24 Bladder Cancer Cells

目的 探讨CXCR4基因沉默后对人膀胱癌T24细胞增殖、凋亡、迁移能力的影响。 方法 将T24细胞分为3组,即干扰组、阴性对照组和空白对照组。采用阳离子脂质体转染试剂Lipofectamine2000将靶向沉默CXCR4基因的小干扰RNA(siRNA)序列转染至膀胱癌T24细胞株中,Western-blot检测转染后T24细胞中CXCR4蛋白的表达水平,MTT比色法检测细胞凋亡,Transwell迁移试验观察T24细胞迁移能力。 结果 转染siRNA后,CXCR4蛋白表达量明显下调(P<0.05),与空白对照组及阴性对照组的T24细胞比较,沉默CXCR4基因后,显著抑制膀胱癌T24细胞的增殖活性(均为P<0.05),促进T24细胞凋亡(均为P<0.05);Transwell实验结果提示,RNAi干扰CXCR4表达后能显著抑制膀胱癌T24细胞的迁移能力(P<0.05)。 结论 RNAi干扰CXCR4表达水平能够抑制T24细胞增殖、促进细胞凋亡及降低细胞的迁移能力,推测CXCR4基因可能是膀胱肿瘤细胞增殖及迁移的重要调控因子之一。

Objective To explore the effect of siRNA targeting the CXCR4 gene on cell proliferation, apoptosis and migration of T24 cells. Methods T24 cells were divided into three groups: interference group, negative control group, and blank control group. Small interfering RNA (siRNA) was transfected together with Lipofectamine2000 in T24 human bladder cancer cells to block the CXCR4 signal pathway. Western-blot analysis was used to confirm the successful suppression of CXCR4 protein by siRNA . The effect of siRNA on cell death, proliferation and invasiveness was assessed by MTT colorimetric method, cell counting kit-8 assay;the effects of apoptosis was detected by FCM;and Transwell migration assay was performed. Results Expression of CXCR4 protein was successfully suppressed after transfected with siRNA which was verified by Western-blot (P<0.05). Compared with blank control and negative control, the proliferation of T24 bladder cancer cells was successfully restrained and the cell death of T24 bladder cancer cells was significantly promoted by inhibition of CXCR4 gene (all P value <0.05 ). Transwell test result demonstrated that migration of T24 bladder cancer cells was significantly inhibited after CXCR4 gene was interfered by siRNA (P<0.05). Conclusion This study showed interfering CXCR4 expression level can inhibit cell proliferation, promote cell apoptosis and decreased cell migration ability, which suggested that CXCR4 gene was an important enhancer for the proliferation and migration of bladder cancer cells.