Zhang Chunhe, Fu Shaowei, Wang Luyue, Wang Fang, Wu Dan, Zhe Xiangyi, Xin Huizhen, Li Hongtao, Li Dongmei, Jin Fuyuan, Shao Renfu, Pan Zemin
Department of Biochemistry and Molecular Biology, School of Medicine, Shihezi University, Xinjiang Endemic and Ethnic Disease and Education Ministry Key Laboratory, Shihezi, Xinjiang 832002, People's Republic of China.
School of Science, Technology and Engineering, Genecology Research Centre, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
Onco Targets Ther. 2021 Jan 14;14:403-411. doi: 10.2147/OTT.S277445. eCollection 2021.
The aim of this study was to determine whether gene methylation and tissue protein expression can be used as a tool with high sensitivity and specificity for cervical cancer screening. We analyzed the correlation between promoter methylation of gene and cervical cancer and high risk HPV16/18 infection.
Tissue samples of normal cervical or chronic cervicitis (n=51), CIN (cervical intraepithelial neoplasia) (n=35), and cervical carcinoma (n=68) were tested for HPV16/18 infection by polymerase chain reaction (PCR). We also detected the methylation status of the gene promoter in the same tissues by methylation-specific PCR (MSP), then analyzed the correlation between promoter methylation and HPV16/18 infection. Immunohistochemistry was used to analyze gene expression in 152 cervical tissues. We detected mRNA expression in cervical tissues (including cancer and non-cancer) by real-time fluorescence quantitative PCR (qPCR).
Among 93 high-grade cervical lesions (CINII and above) and cervical cancer samples, 57 cases were positive for HPV16/18 infection and 36 cases were negative. gene methylation occurred in 9 out of 51 cases in normal cervical tissues (17.6%), 16 of 35 cases in CIN tissues (45.7%), and 50 of 68 cases in cervical cancer (73.5%). The differences in methylation rate of the three groups were statistically significant (<0.05). The methylation rate in the positive HPV16/18 infection group was 73.7%, while the negative group was 63.9%. Compared with normal tissues, ZNF582 protein was highly expressed in cervical cancer tissues, but mRNA expression was low.
While ZNF582 protein is highly expressed in cervical cancer tissues, it was not sufficient for use as a standard for cervical cancer staging. On the other hand, promoter methylation had high specificity and sensitivity in detecting CINII and highly diseased cervical lesions and could be used as a diagnostic marker for cervical cancer of women.
本研究的目的是确定基因甲基化和组织蛋白表达是否可作为一种具有高灵敏度和特异性的宫颈癌筛查工具。我们分析了基因启动子甲基化与宫颈癌及高危型HPV16/18感染之间的相关性。
采用聚合酶链反应（PCR）检测正常宫颈或慢性宫颈炎组织样本（n = 51）、宫颈上皮内瘤变（CIN）组织样本（n = 35）和宫颈癌组织样本（n = 68）中的HPV16/18感染情况。我们还通过甲基化特异性PCR（MSP）检测相同组织中基因启动子的甲基化状态，然后分析启动子甲基化与HPV16/18感染之间的相关性。采用免疫组织化学方法分析152例宫颈组织中的基因表达情况。通过实时荧光定量PCR（qPCR）检测宫颈组织（包括癌组织和非癌组织）中的mRNA表达情况。
在93例高级别宫颈病变（CINII及以上）和宫颈癌样本中，57例HPV16/18感染呈阳性，36例呈阴性。正常宫颈组织51例中有9例发生基因甲基化（17.6%），CIN组织35例中有16例发生甲基化（45.7%），宫颈癌组织68例中有50例发生甲基化（73.5%）。三组甲基化率差异具有统计学意义（<0.05）。HPV16/18感染阳性组的甲基化率为73.7%，阴性组为63.9%。与正常组织相比，ZNF582蛋白在宫颈癌组织中高表达，但mRNA表达较低。
虽然ZNF582蛋白在宫颈癌组织中高表达，但不足以作为宫颈癌分期的标准。另一方面，启动子甲基化在检测CINII及高度病变的宫颈病变方面具有高特异性和敏感性，可作为女性宫颈癌的诊断标志物。
