宫颈癌中高危型HPV感染与miR-362-3p、Nemo样激酶表达的相关性研究  

Correlation Between High-risk HPV Infection and Expression of miR-362-3p and Nemo-like Kinase in Cervical Cancer

作　　者：赵虹 张芬 张振东 ZHAO Hong;ZHANG Fen;ZHANG Zhendong(The Second Affiliated Hospital of Zunyi Medical University,Guizhou 563000,China;Affiliated Hospital of Zunyi Medical University,Guizhou 563000,China)

机构地区：[1]遵义医科大学第二附属医院,遵义563000 [2]遵义医科大学附属医院,遵义563000

出　　处：《病毒学报》2021年第6期1363-1369,共7页Chinese Journal of Virology

摘　　要：高危型人乳头瘤病毒(High-risk human papillomavirus,HPV)感染是宫颈癌发病的重要因素,但高危型HPV感染引起宫颈上皮恶变的机制尚不清楚。微小RNA(micro RNA,miR)-362-3p是具有抑癌活性的miR,在宫颈癌中表达降低;Nemo样激酶(Nemo-like kinase,NLK)是生物信息学预测得到的miR-362-3p靶基因,在宫颈癌中表达增加。但宫颈癌发病过程中高危型HPV感染与miR-362-3p、NLK的关系尚不清楚。本研究检测了miR-362-3p、NLK在宫颈癌组织及宫颈癌细胞株中的变化,并通过转染miR-362-3p模拟物、NLK表达质粒的方式验证了miR-362-3p靶向NLK调节高危型HPV阳性宫颈癌细胞增殖的作用。结果显示:与癌旁组织比较,宫颈癌组织中miR-362-3p的表达明显降低、NLK的表达明显增加且与高危型HPV阴性的宫颈癌组织比较,高危型HPV阳性的宫颈癌组织中miR-362-3p的表达明显降低、NLK的表达明显增加;与正常宫颈上皮细胞比较,HPV16感染的SiHa细胞、HPV18感染的HeLa细胞及HPV阴性的C33A细胞中miR-362-3p的表达明显降低、NLK的表达明显增加且SiHa细胞中miR-362-3p表达降低、NLK表达增加最明显;在SiHa细胞中,过表达miR-362-3p能够降低细胞活力及NLK的表达,同时也使NLK双荧光素酶报告基因的荧光活力降低;过表达NLK能够逆转miR-362-3p降低细胞活力及NLK表达的作用。以上结果表明宫颈癌中高危型HPV感染与miR-362-3p低表达、NLK高表达有关,过表达miR-362-3p通过靶向抑制NLK降低高危型HPV感染宫颈癌细胞的增殖活力。本研究的创新点在于初步探索了高危型HPV感染促进宫颈癌细胞增殖的作用及机制,在高危型HPV感染的宫颈癌中miR-362-3p表达减少并靶向引起NLK表达增加,进而促进了宫颈癌细胞的增殖,这为将来深入认识高危型HPV引起宫颈癌发病的机制提供依据。High-risk human papillomavirus(HPV)infection is an important factor in the pathogenesis of cervical cancer. However,the mechanism of cervical epithelial malignant transformation caused by high - risk HPV infection is not clear. MicroRNA-362-3 p(miR-362-3 p)has antitumor activity,and its expression is reduced in cervical cancer. Nemo-like kinase(NLK)is a target gene of miR-362-3 p according to bioinformatics analysis,and its expression is increased in cervical cancer. However,the relationship between high -risk HPV infection and miR-362-3 p and NLK is not clear. Here,changes in the expression of miR-362-3 p and NLK in cervical-cancer tissue and cervical-cancer cell lines were measured. Also,the effect of miR-362-3 p on the proliferation of high -risk HPV -positive cervical cancer cells by targeting NLK was verified by transfection of a miR - 362 -3 p mimic and NLK expression plasmid. We discovered that,compared with paracancerous tissue,miR - 362 - 3 p expression was decreased significantly and that NLK expression was increased significantly in cervical cancer.Also,compared with high-risk HPV-negative cervical cancer,miR-362-3 p expression was reduced significantly and NLK expression increased significantly in high-risk HPV -positive cervical cancer. Compared with normal cervical epithelial cells,miR - 362 - 3 p expression was decreased significantly and NLK expression increased significantly in HPV16-infected SiHa cells,HPV18-infected HeLa cells and HPV-negative C33 A cells. miR-362-3 p overexpression decreased cell viability and NLK expression,whereas mir-362-3 p overexpression also decreased the mean fluorescence intensity of the NLK double - luciferase reporter gene. NLK overexpression reversed the decreasing effect of miR-362-3 p on cell viability and NLK expression. These results suggest that high -risk HPV infection in cervical cancer is related to low expression of miR - 362 -3 p and high expression of NLK. mir-362-3 p overexpression can reduce the proliferation of high-risk HPV-infected cervical cancer ce

关 键 词：宫颈癌 高危型HPV miR-362-3p Nemo样激酶 增殖 

分 类 号：R373.9[医药卫生—病原生物学]