晚期糖基化终末产物受体对宫颈癌细胞生物学行为的影响

Effect of receptor for advanced-glycation end products on proliferation, apoptosis and tumor formation of cervical cancer Si Ha cells in nude mice

目的探讨晚期糖基化终末产物受体(RAGE)对宫颈癌SiHa细胞增殖、凋亡等生物学行为及裸鼠成瘤能力的影响及相关机制。方法 SiHa细胞分3组培养,空白对照组为仅添加凝聚胺,空载体组为添加凝聚胺+转染空载体质粒,过表达组为添加凝聚胺+转染RAGE过表达质粒。采用p Lenti-C-m GFP-RAGE质粒转染方法构建RAGE基因稳定过表达的宫颈鳞癌SiHa细胞。采用Western blot法检测SiHa细胞RAGE、增殖细胞核抗原(PCNA)、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、与Bcl-2相互作用的细胞死亡调节因子(Bim)蛋白表达水平,CCK-8法检测RAGE基因对SiHa细胞增殖的影响,流式细胞仪检测RAGE基因对SiHa细胞凋亡的影响。采用裸鼠皮下成瘤法,动态观察RAGE基因对宫颈癌肿瘤生长的影响。结果转染p Lenti-C-m GFP-RAGE质粒可明显上调SiHa细胞中RAGE蛋白的表达。过表达组SiHa细胞的增殖力均明显高于空白对照组及空载体组(均P<0.05),而过表达组SiHa细胞的凋亡率均明显低于空白对照组及空载体组(均P<0.05)。过表达组PCNA及Bcl-2蛋白表达水平均明显高于空白对照组及空载体组(均P<0.05),而Bax/Bcl-2表达水平均明显低于空白对照组及空载体组(均P<0.05),3组Bax及Bim蛋白表达水平比较差异均无统计学意义(均P>0.05)。过表达组裸鼠皮下瘤体体积和瘤体质量均显著高于空载体组的体积和瘤体质量,差异均有统计学意义(均P<0.05)。结论上调RAGE基因的表达可促进SiHa细胞增殖,抑制SiHa细胞凋亡,促进宫颈癌瘤体的生长,该作用和抗凋亡蛋白Bcl-2相关。

Objective To investigate the effects and mechanisms of receptor for advanced-glycation end products（RAGE） on the proliferation, apoptosis and tumor growth of cervical cancer SiHa cells in nude mice. Methods The culturedSiHa cells were divided into three groups： the blank control group was only treated with polyamines, the vector group wastreated with polyamines and transfected with empty vector plasmid, and the RAGE group was treated with polyamines andtransfected with plasmid pLenti-C-mGFP-RAGE. The expression of RAGE protein, proliferation-related protein PCNA andapoptotic-related proteins Bcl-2, Bax and Bim were detected by Western blot. The proliferation and apoptosis of SiHa cellswere examined with CCK-8 and flow cytometry, respectively. The SiHa cells in3 groups were inoculated subcutaneously innude mice and the growth of transplanted tumor was measured dynamically and compared in 3 groups. Results ThepLenti-C-mGFP-RAGE transfection significantly increased RAGE expression in SiHa cells. The cell proliferation of RAGE groupwas significantly higher than that of blank control group and vector group（both P〈0.05）, while the cell apoptosis of RAGE groupwas significantly lower than that of blank control group and vector group（both P〈0.05）. The expression of PCNA and Bcl-2proteins in RAGE group was significantly higher than that in blank control group and vector group（both P〈0.05）, while theexpression of Bax/Bcl-2 protein significantly reduced （both P〈0.05）, but there was no significant difference in expression of Baxand Bim proteins. The tumor size and weight in RAGE group [（1 599.47±247.97）mm3, （1.67±0.35）g] were significantly greaterthan those in vector group [（629.92±140.59）mm3, （0.74±0.42）g, both P〈0.05]. Conclusion Up-regulation of RAGE canenhance the proliferation of SiHa cells, inhibit cell apoptosis, and promote the growth of cervical cancer in nude mice, which are associated with the regulation of apoptosis-related protein expressions.