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目的:探讨连接蛋白43(connexin 43,Cx43)对膀胱癌细胞侵袭能力的影响及其可能的作用机制。方法:选取2014年6月至2015年9月间承德医学院附属医院泌尿外科52例膀胱癌手术组织标本及32例癌旁组织,以及人膀胱癌细胞株5637。用免疫组化方法检测膀胱癌组织中Cx43蛋白表达。将Cx43脂质体、空白脂质体、siRNA及siRNA对照质粒转染5637细胞,Western blotting验证过Cx43表达和干扰效果;用Transwell侵袭实验检测5637细胞侵袭能力的变化,用Western blotting检测5637细胞MMP-2、MMP-9和P-P38蛋白表达水平的变化。结果:膀胱癌组织中Cx43蛋白的表达水平明显高于癌旁组织[(5.21±0.33)vs(2.84±0.19),P<0.01]。转染Cx43脂质体和siRNA成功上调/下调5637细胞中Cx43的表达。Cx43过表达组5637细胞的侵袭能力高于对照组[穿膜细胞数:(1.36±0.04)vs(0.70±0.15)个,P<0.01],siRNA干扰组细胞的侵袭能力低于对照组[穿膜细胞数:(0.20±0.08)vs(0.59±0.13)个,P<0.05)。Cx43过表达组细胞MMP-2、MMP-9、P-P38/P38蛋白水平均高于对照组(P<0.01或P<0.05),siRNA干扰组均低于对照组低(均P<0.01)。结论:Cx43增强膀胱癌5637细胞的侵袭能力,其机制可能是通过激活P38/MAPK信号途径实现的。
Objective: To investigate the effect of connexin 43 (Cx43) on the invasiveness of bladder cancer cells and its possible mechanism. Methods: Twenty-two cases of bladder cancer and 32 cases of bladder cancer tissues and 5637 human bladder cancer cell lines were selected from Department of Urology, Affiliated Hospital of Chengde Medical College from June 2014 to September 2015. The expression of Cx43 in bladder cancer tissues was detected by immunohistochemistry. The Cx43 liposomes, blank liposomes, siRNA and siRNA control plasmid were transfected into 5637 cells, and the expression of Cx43 and the interference effect were verified by Western blotting. The invasion ability of 5637 cells was detected by Transwell invasion assay. The expression of 5637 cells was detected by Western blotting -2, MMP-9 and P-P38 protein expression levels. Results: The expression of Cx43 protein in bladder cancer tissues was significantly higher than that in adjacent tissues [(5.21 ± 0.33) vs (2.84 ± 0.19), P <0.01]. Transfection with Cx43 liposomes and siRNA successfully up-regulated / down-regulated Cx43 expression in 5637 cells. The invasion ability of 5637 cells in Cx43 overexpression group was higher than that in control group [(1.36 ± 0.04) vs (0.70 ± 0.15), P <0.01]. The invasion ability of siRNA interference group was lower than that of control group Membrane cells: (0.20 ± 0.08) vs (0.59 ± 0.13), P <0.05). The protein levels of MMP-2, MMP-9 and P-P38 / P38 in Cx43 overexpression group were significantly higher than those in control group (P <0.01 or P <0.05) . Conclusion: Cx43 enhances the invasiveness of bladder cancer 5637 cells through its activation of P38 / MAPK signaling pathway.