Survivin通过ERK/E-cadherin信号通路促进子宫内膜癌细胞增殖和迁移

Survivin promotes proliferation and migration of endometrial carcinoma cells via ERK/E-cadherin signaling pathway

目的 探讨Survivin在子宫内膜癌发生发展中的作用及其与ERK/E-cadherin信号通路的关系.方法 免疫组织化学方法检测Survivin在子宫内膜癌及癌旁组织中的表达情况,将含有Survivin特异性siRNA的重组质粒p RNAT-suv转染子宫内膜癌细胞株KLE,RT-PCR检测转染前后各组细胞Survivin基因mRNA表达水平的变化,CCK-8法检测Survivin干扰表达后细胞增殖能力的变化, Transwell法检测Survivin干扰表达后细胞迁移能力的变化,Western blotting法检测转染前后各组细胞β-catenin、cyclinD1、c-myc和p-ERK、Slug、E-cadherin蛋白表达差异.结果 Survivin在子宫内膜癌组织中的表达明显高于癌旁组织(P<0.05),且低分化癌Survivin表达高于高、中分化癌(P=0.03),深肌层浸润癌高于浅肌层浸润癌(P=0.04).Survivin干扰表达后细胞增殖和迁移能力均发生明显的下降(P<0.01),Western blotting结果显示Survivin干扰表达后,β-catenin及其下游增殖相关分子cyclinD1、c-myc等表达都明显下调,p-ERK、Slug等转移相关蛋白发生明显的表达下调,E-cadherin表达上调.结论 Survivin的高表达与子宫内膜癌的发生发展密切相关,信号通路研究发现Survivin可能主要是通过ERK/E-cadherin通路来促进子宫内膜癌细胞的增殖和转移.

Objective To investigate the effect of Survivin in the occurrence and development of endometrial carcinoma and its relationship with ERK / E - cadherin signaling pathway. Methods Immunohistochemistry was used to examine the expression of Survivin in endometrial carcinoma and adjacent tissues. Recombinant plasmid pRNAT - suv containing Survivin - specific siRNA was transfected into endometrial carcinoma cell line KLE. The mRNA expression level of Survivin was determined by RT-PCR before and after the transfection. CCK-8 method was used to examine the cell proliferation after Survivin interference on the expression. Transwell method was used to examine the cell migration after Survivin interference on the expression. Western blotting assay was used to examine the differences in the protein expression of β-catenin,cyclin D1,c-myc,p-ERK,Slug and E-cadherin between the groups before and after transfection. Results The expression level of Survivin in the endometrial carcinoma tissues was significantly higher than that in the adjacent tissues(P<0.05). The expression level of Survivin in poorly differentiated endometrial carcinoma was higher than that in the well and moderately differentiated endometrial carcinoma (P=0.03). The expression level of Survivin in deep myometrial invasive carcinoma was higher than that in superficial myometrial invasive carcinoma(P=0.04). After Survivin interference on the expression,the cell proliferation and migration significantly decreased(P<0.01). The findings of Western blotting results showed that after Survivin interference,the expression levels of β-catenin and its downstream proliferation-related molecules cyclin D1 and c-myc were down-regulated,the expression levels of p-ERK,Slug and metastasis-related proteins were significantly down-regulated,and the expression level of E-cadherin was up-regulated. Conclusion The high expression of Survivin is closely related to the occurrence and development of endometrial carcinoma. This signaling pathway study revealed that Survivin may promote the proliferation and metastasis of endometrial carcinoma cells mainly via ERK/E-cadherin pathway.