HPV16阳性和阴性患者宫颈癌组织中miR-27a-3p和HOXB8蛋白的表达差异及其机制研究

发布时间：2018-04-23 20:27

本文选题：人乳头瘤病毒型 + 宫颈癌　；参考：《中国药房》2017年23期

【摘要】：目的:探讨mi R-27a-3p和同源盒B8(HOXB8)蛋白在人乳头瘤病毒16型阳性[HPV16(+)]和阴性[HPV16(-)]患者宫颈癌组织中表达的差异及其机制。方法:选取2012年1月-2016年1月于我院就诊的宫颈癌患者120例,根据其是否感染HPV16分为HPV16(+)组(60例)和HPV16(-)组(60例)。采用实时荧光定量聚合酶链反应法检测两组患者宫颈癌组织中mi R-27a-3p和HOXB8 m RNA的表达水平,采用蛋白印迹法检测HOXB8蛋白的表达水平,采用巢式降落式甲基化特异性聚合酶链反应法检测mi R-27a-3p启动子区DNA甲基化水平,采用染色质免疫共沉淀-定量聚合酶链反应法检测mi R-27a-3p启动子区组蛋白甲基化水平;培养人宫颈癌细胞系Si Ha细胞株,考察转染mi R-27a-3p模拟物(mimic)和抑制物(inhibitor)后mi R-27a-3p和HOXB8 m RNA的表达水平。结果:HPV16(+)组患者宫颈癌组织中mi R-27a-3p的表达水平显著低于HPV16(-)组患者,HOXB8 m RNA和蛋白表达水平、mi R-27a-3p启动子区DNA甲基化水平和组蛋白甲基化水平均显著高于HPV16(-)组患者,差异均有统计学意义(P0.05或P0.01)。在Si Ha细胞转染mi R-27a-3p mimic后,mi R-27a-3p的表达水平显著升高,而HOXB8 m RNA的表达水平显著降低;转染mi R-27a-3p inhibitor后,mi R-27a-3p的表达水平显著降低,而HOXB8 m RNA的表达水平则显著升高,差异均有统计学意义(P0.01)。结论:HPV16可能通过启动子区DNA甲基化和组蛋白甲基化下调mi R-27a-3p的表达,从而影响宫颈癌的发生与发展,HOXB8蛋白可能是其作用靶点。
[Abstract]:Objective: to investigate the expression of mi R-27a-3p and homologous box B8 (HOXB8) protein in human papillomavirus 16 positive [HPV16 ()] and negative [HPV16 ()] cervical cancer tissues and its mechanism. Methods: 120 patients with cervical cancer from January 2012 to January 2016 were divided into two groups: HPV16 (60 cases) and HPV16- (60 cases) according to whether they were infected or not. The expression of mi R-27a-3p and HOXB8 m RNA in cervical cancer tissues was detected by real-time fluorescence quantitative polymerase chain reaction, and the expression of HOXB8 protein was detected by Western blot. The DNA methylation level in the mi R-27a-3p promoter was detected by nested landing methylation specific polymerase chain reaction (NPCR), and histone methylation in the mi R-27a-3p promoter was detected by chromatin immunoprecipitation and quantitative polymerase chain reaction (QPCR). The expression of mi R-27a-3p and HOXB8 m RNA in human cervical cancer cell line SIHA was studied after transfection of mi R-27a-3p mimicoid and inhibitor. Results the expression level of mi R-27a-3p in cervical cancer tissues of patients with HPV16 (10% HPV16) was significantly lower than that of patients with HPV16-). The levels of DNA methylation and histone methylation in the promoter region of mi R-27a-3p were significantly higher than those in HPV16-) group, and the expression level of HPVB8 m RNA and protein in HPV16- group were significantly higher than those in HPV16- group. The difference was statistically significant (P 0.05 or P 0.01). After transfection of mi R-27a-3p mimic into SIHA cells, the expression level of mi R-27a-3p was significantly increased, while the expression of HOXB8 m RNA was significantly decreased, and the expression of HOXB8 m RNA was significantly decreased after transfection of mi R-27a-3p inhibitor, while that of HOXB8 m RNA was significantly increased. The difference was statistically significant (P 0.01). Conclusion the expression of mi R-27a-3p may be down-regulated by DNA methylation and histone methylation in the promoter region of HPV16, which may be a possible target for carcinogenesis and development of cervical cancer.
【作者单位】：牡丹江医学院附属红旗医院妇产科;
【分类号】：R737.33

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