NLRC5抑制自噬促进子宫内膜癌细胞增殖的研究

NLRC5 promotes proliferation of endometrial cancer cells by inhibiting autophagy

目的初步探究NLRC5对子宫内膜癌细胞AN3CA增殖的作用及其机制。方法选取20例子宫内膜癌患者手术切除的子宫内膜腺癌标本及门诊病理显示正常的子宫内膜标本20例,免疫组化实验检测子宫内膜癌组织和正常子宫内膜组织标本中NLRC5及自噬相关蛋白Beclin1、LC3的表达。Western blot及qRT-PCR法检测子宫内膜癌组织和正常子宫内膜组织标本及子宫内膜癌AN3CA细胞中NLRC5及自噬相关蛋白Beclin1、LC3的表达。CCK-8法检测AN3CA细胞增殖。结果子宫内膜癌组与正常子宫内膜组相比,NLRC5蛋白表达下调,自噬相关蛋白Beclin1、LC3表达上调(P <0. 05);转染NLRC5质粒后,转染组与未转染组相比,自噬蛋白Beclin1、LC3的蛋白表达水平显著下调(P <0. 05);转染NLRC5质粒后,转染组与未转染组相比,转染组细胞增殖明显增加(P <0. 05);转染NLRC5质粒后并加自噬激动剂雷帕霉素,结果显示,转染NLRC5质粒组与未转染NLRC5质粒组相比,细胞增殖明显增加(P <0. 05);转染NLRC5质粒后同时加雷帕霉素组与只转染NLRC5质粒组相比,细胞自噬水平升高,但细胞增殖明显下降(P <0. 05)。结论 NLRC5可能通过抑制自噬进而促进子宫内膜癌细胞增殖。

Objective To preliminarily explore the effect and mechanism of NLRC5 on the proliferation of endometrial cancer cells AN3 CA. Methods We selected 20 specimens of endometrial adenocarcinoma surgically removed from patients with endometrial cancer and 20 cases of normal endometrial specimens in outpatient pathology.The protein expression of NLRC5 and autophagy-related proteins Beclin1, LC3 in endometrial cancer tissue and normal endometrial tissue specimens were observed by immunohistochemistry. Expression of NLRC5 and autophagy-related proteins Beclin1, LC3 in endometrial carcinoma and normal endometrial tissue specimens and endometrial carcinoma AN3 CA cells were detected by Western blot and qRT-PCR. Proliferation of AN3 CA cells was detected by the method of cck-8. Results The expression of NLRC5 protein was down-regulated and the expression of autophagy-related proteins Beclin1, LC3 were up-regulated in endometrial carcinoma tissues compared with normal endometrial tissues(P<0.05). After transfection of NLRC5 plasmid, the expression levels of autophagy-related proteins Beclin1 and LC3 were significantly down-regulated in the transfected group compared with the untransfected group(P<0.05). After transfection of NLRC5 plasmid, the cell proliferation of the transfected group was significantly increased compared with the untransfected group(P<0.05). After transfecting the NLRC5 plasmid and treating with rapamycin, cell proliferation was significantly increased in the transfected NLRC5 plasmid group compared with the untransfected NLRC5 plasmid group(P<0.05). Compared with NLRC5 plasmid group, the autophagy level was increased, but the cell proliferation was significantly decreased(P<0.05). Conclusion NLRC5 may promote the proliferation of human endometrial cancer cells by inhibiting autophagy.