干扰水通道蛋白3对胃癌细胞增殖与凋亡的影响

Effects of inhibiting aquaporin-3 expression on the proliferation and apoptosis of gastric cancer cells

目的:探究水通道蛋白3(AQP3)对胃癌细胞增殖与凋亡的影响及其可能机制。方法:通过RT-PCR、Western-blot实验检测胃癌细胞株中的AQP3表达水平;小干扰RNA构建AQP3低表达细胞系;CCK-8增殖实验和流式细胞术分别检测细胞增殖与凋亡水平;Western-blot检测细胞自噬水平,透射电镜观察自噬小体数目。结果:胃癌细胞AQP3表达水平较高;与干扰前相比,AQP3低表达细胞系中胃癌细胞增殖水平降低(P<0.05),凋亡水平提高(P<0.001),自噬标记蛋白LC3Ⅱ水平降低(P<0.001),P62表达水平上升(P<0.001)。结论:AQP3在胃癌细胞中高表达,干扰AQP3可通过降低自噬水平促进细胞凋亡,进而抑制胃癌细胞增殖,提示AQP3具有成为胃癌治疗分子靶点的潜能。

Objective:To investigate the effects of inhibiting aquaporin3(AQP3)on the proliferation and apoptosis of gastric cancer cells and the potential mechanisms.Methods:The expression level of AQP3in gastric cancer cell lines was determined by RT PCR and Western blot.AQP3low expression cell line was constructed by small interfering RNA.CCK8proliferation assay and flow cytometry were used to detect the cell proliferation and apoptosis.The autophagy level was detected by Western blot and number of autophagosomes was measured by transmission electron microscopy.Results:Significantly higher expression of AQP3was seen in the gastric cancer cells,yet was markedly down regulated following interference(P<0.05).Increased apoptotic level,P62expression and decreased autophagy biomarker of LC3Ⅱwere also seen in the gastric cancer cells after interfering(all P<0.001).Conclusion:AQP3is highly expressed in gastric cancer cells,and interfering the expression of AQP3can promote cell apoptosis by decreasing the level of autophagy and inhibiting the proliferation of gastric cancer cells.This suggests that AQP3may be potential target for treating gastric cancer.