RNAi抑制galectin-3基因表达对宫颈癌细胞增殖及凋亡的影响机制

Effect and mechanism of silencing galectin-3 gene expression by RNA interference on the proliferation and apoptosis of cervical carcinoma cell line

目的探讨RNAi抑制半乳糖凝集素(galectin-3)基因表达对宫颈癌细胞增殖及凋亡的影响及机制。方法提取宫颈癌组织和相应的癌旁组织中的总蛋白,Western blot检测galectin-3的蛋白表达;人宫颈癌He La细胞分为正常对照组、siRNA-control组和siRNA-galectin-3组,各组细胞转染48 h后,Western blot检测galectin-3的蛋白表达;CCK8实验和流式细胞仪分别检测细胞的增殖及凋亡情况;Western blot检测ki67、PCNA、cleaved caspase3、β-catenin、cyclin D1蛋白表达。结果 galectin-3在宫颈癌组织中的蛋白表达显著高于癌旁组织(P<0.01);转染siRNA-galectin-3能显著降低galectin-3的蛋白表达;与正常对照组和siRNA-control组比较,siRNA-galectin-3组细胞存活率显著降低,细胞凋亡率显著升高,ki67、PCNA阳性细胞数及ki67、PCNA、β-catenin、cyclin D1蛋白表达显著降低,cleaved caspase3蛋白显著上调表达(P<0.01)。结论 RNAi抑制galectin-3基因表达可降低宫颈癌细胞的增殖,诱导细胞的凋亡,其机制与抑制Wnt/β-catenin信号通路有关。

Objective To explore the effect and mechanism of silencing galectin-3 gene expression by RNA interference on the proliferation and apoptosis of cervical carcinoma cell line. Methods The total protein in the cervical cancer tissues and the cor- responding cancer-adjacent tissues was extracted, and then the expression of galectin-3 protein was detected by Western blot. Hu- man cervical carcinoma HeLa cells were divided into the normal control group, the siRNA-control group and the siRNA-galectin- 3 group, and the cells in each group were transfeeted for 48 hours, then the expression of galectin-3 protein was detected by Western blot. Cell proliferation and cell apoptosis were respectively detected by CCK8 assay and flow cytometry. The expression of ki67, proliferating cell nuclear antigen （PCNA） , cleaved caspase3, β-catenin and cyclin D1 proteins were detected by Western blot. Results The expression of galectin-3 protein was significantly higher in the cervical carcinoma tissues than in the paracancerous tissues （ P〈0.01 ）. siRNA-galectin-3 transfection could significantly reduce the expression of galectin-3 protein. The cell survival rate in the siRNA-galectin-3 group decreased significantly as compared with the normal control group and the siRNA-control group, while the cell apoptosis rate increased significantly. The number of positive cells of ki67 and PCNA and the expression of ki67, PCNA, β-catenin and cyclin D1 proteins in the siRNA-galectin-3 group significantly reduced as compared with the normal control group and the siRNA-control group, while the expression of cleaved caspase3 was significantly up-regulated （P〈0.01 ）. Conclusions Inhibition of galectin-3 gene expression by RNA interference can reduce the proliferation of cervical cancer cells and induce cell apoptosis, and the mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.