hnRNP E1与HPV16早期基因E2和E6在宫颈癌变中的作用及交互效应

Effects of hnRNP E1 and both early genes E2 and E6 of HPV16 together with their interactions on cervical carcinogenesis

目的探讨核不均一核糖核蛋白(hnRNP)E1与HPV16早期基因E2和E6在宫颈癌变进展中的作用及相互关系。方法从在山西省介休市建立的自然人群宫颈病变队列中,选取2014年6-9月经病理学确诊的正常宫颈(NC)女性56例,低度宫颈上皮内瘤变(CIN Ⅰ)病例58例和高度宫颈上皮内瘤变(CIN Ⅱ/Ⅲ)病例50例,以及同期在山西省肿瘤医院确诊的40例宫颈鳞状细胞癌(SCC)病例作为研究对象。采用结构式问卷收集研究对象的宫颈病变相关资料的同时,采集宫颈组织活检标本和宫颈脱落细胞。采用导流杂交法检测HPV感染状况,采用Western Blot检测hnRNP E1以及HPV16 E2和E6蛋白表达水平。采用SPSS 22.0软件进行Kruskal-Wallis H检验、χ^2检验、趋势χ^2检验和logistic回归分析,采用广义多因子降维法(GMDR)评价交互作用。结果HPV16感染率在CINⅠ(15.52%,9/58)、CINⅡ/Ⅲ(40.00%,20/50)和SCC组(67.50%,27/40)均高于NC组(8.93%,5/56),并随着宫颈病变程度的加重呈现升高趋势(趋势检验χ^2=43.613,P<0.001)。hnRNP E1表达量在不同宫颈病变组间差异有统计学意义(H=9.98,P=0.019),且随宫颈病变程度的加重呈现降低趋势(趋势检验χ^2=9.495,P=0.002)。HPV16 E2(H=16.20,P=0.001)和E6(H=15.44,P=0.001)表达水平在不同宫颈病变组间差异均有统计学意义。采用GMDR分析显示,hnRNP E1低表达、HPV16 E2低表达与HPV16 E6高表达在CIN Ⅱ/Ⅲ和SCC组中存在交互作用(P<0.05),而在CIN Ⅰ组未发现存在交互效应(P>0.05)。结论hnRNP E1低表达和HPV16早期基因的异常表达可能会增加宫颈病变的发病风险,并在宫颈癌变中存在交互作用。

Objective To explore the effects of hnRNP E1 and both early genes E2 and E6 of HPV16 as well as their interactions in the progression of cervical carcinogenesis. Methods Subjects of this study included 56 women with normal cervix (NC), 58 patients with low-grade cervical intraepithelial neoplasm (CINⅠ) and 50 patients with high-grade cervical intraepithelial neoplasm (CINⅡ/Ⅲ) who were all recruited from the 'Cervical Lesions Study Cohort Project’ in Jiexiu of Shanxi province from June to September, 2014. Another 40 patients with cervical squamous cell carcinoma (SCC) were from the Shanxi Tumor Hospital during the same period. Information related to cervical lesions were collected, using a structured questionnaire, with cervical tissues and cervical exfoliated cells gathered from all the participants. HPV infection was detected by flow-through hybridization, while the levels of expression on hnRNP E1, HPV16 E2 and E6 protein were measured by Western Blot. Kruskal-Wallis H test,χ^2 test, trend χ^2 test were analyzed by SPSS 22.0 software, while interaction was evaluated by generalized multifactor dimensionality reduction (GMDR). Results The overall infection rates of HPV16 related to CINⅠ(15.52%, 9/58), CINⅡ/Ⅲ(40.00%, 20/50) and SCC (67.50%, 27/40) groups were all higher than that of the NC group (8.93%, 5/56) and with an increasing trend on the severity of cervical lesions (trend χ^2=43.613, P<0.001). The levels of expression on hnRNP E1 protein were significantly different in the groups with different cervical lesions (H=9.98, P=0.019), showing a decreasing trend with the severity of cervical lesions (trend χ^2=9.495, P=0.002). The levels of expression on HPV16 E2 (H=16.20, P=0.001) and HPV16 E6 (H=15.44, P=0.001) were significantly different in groups with different cervical lesions. Results of GMDR showed that the best interaction model in both groups of CINⅡ/Ⅲ and SCC appeared as hnRNP E1 low expression, HPV16 E2 low expression and HPV16 E6 high expression. However, no similar interaction was seen in CINⅠ(P>0.05). Conclusions Both low expressions of hnRNP E1 and abnormal expression of HPV16 E2 and E6 could increase the risk of high-grade CIN and cervical cancer. It seemed that they might have an important synergistic effect on the progression of cervical cancer.