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-中国药房

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

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篇名：	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
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摘要：	目的：探讨miR-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miR-27a-3p和HOXB8 mRNA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miR-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miR-27a-3p启动子区组蛋白甲基化水平；培养人宫颈癌细胞系SiHa细胞株，考察转染miR-27a-3p模拟物（mimic）和抑制物（inhibitor）后miR-27a-3p和HOXB8 mRNA的表达水平。结果：HPV16（+）组患者宫颈癌组织中miR-27a-3p的表达水平显著低于HPV16（－）组患者，HOXB8 mRNA和蛋白表达水平、miR-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（P＜0.05或P＜0.01）。在SiHa细胞转染miR-27a-3p mimic后，miR-27a-3p的表达水平显著升高，而HOXB8 mRNA的表达水平显著降低；转染miR-27a-3p inhibitor后，miR-27a-3p的表达水平显著降低，而HOXB8 mRNA的表达水平则显著升高，差异均有统计学意义（P＜0.01）。结论：HPV16可能通过启动子区DNA甲基化和组蛋白甲基化下调miR-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 difference and its mechanism of miR-27a-3p and HOXB8 protein in cervical cancer tissues of HPV16-positive and HPV16-negative patients. METHODS： A total of 120 patients with cervical cancer in our hospital during Jan. 2012-Jan. 2016 were divided into HPV16-positive group （60 cases） and HPV16-negative group （60 cases） according to HPV16 infection situation. The expression of miR-27a-3p mRNA and HOXB8 mRNA in cervical cancer tissue were detected by RT-qPCR. The expression of HOXB8 protein was detected by Western blotting assay. DNA methylation level of miR-27a-3p promoter region was detected by nested-down methylation specific PCR （nMS-PCR）. Histone methylation level of miR-27a-3p promoter region was detected by chromatin immunoprecipitation PCR （CHIP-PCR）. The expression of miR-27a-3p mRNA and HOXB8 mRNA were detected after transfecting miR-27a-3p mimic and inhibitor into Human cervical cancer cell line SiHa， respectively. RESULTS： The expression of miR-27a-3p in HPV16-positive group was significantly lower than HPV16-negative group， while HOXB8 mRNA and protein expression， DNA and histone methylation levels of miR-27a-3p promoter region were significantly higher than HPV16-negative group， with statistical significance （P＜0.05 or P＜0.01）. After transfecting miR-27a-3p mimic into SiHa cells， the expression of miR-27a-3p was increased significantly， while that of HOXB8 mRNA was decreased significantly； after miR-27a-3p inhibitor transfection， the expression of miR-27a-3p was decreased significantly， while that of HOXB8 mRNA was increased significantly， with statistical significance （P＜0.01）. CONCLUSIONS： HPV16 may down-regulate the expression of miR-27a-3p through DNA methylation and histone methylation of promoter region， so as to influence the generation and development of cervical cancer. HOXB8 may be the target protein of miR-27a-3p.
期刊：	2017年第28卷第23期
作者：	李会影，王慧智，张子旸
AUTHORS：	LI Huiying，WANG Huizhi，ZHANG Ziyang
关键字：	人乳头瘤病毒16型；宫颈癌；miR-27a-3p；同源盒B8蛋白；DNA甲基化；组蛋白甲基化
KEYWORDS：	Human papilloma virus type 16； Cervical cancer； miR-27a-3p； Homeobox B8 protein； DNA methylation； Histone methylation
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