HPV16/18感染导致宫颈癌的分子机制研究

Molecular mechnism of cervical cancer tumorigenesis correlated with HPV16/18 infection

目的探讨高危型HPV16/18感染对宫颈组织中抑癌基因Rap1GAP的影响,揭示HPV16/18感染导致宫颈癌的分子机制。方法收集因子宫肌瘤手术切除的正常宫颈组织20例和宫颈癌组织20例。检测HPV16/18感染情况后,Trizol法提取组织总RNA,RT-PCR检测组织中Rap1GAP mRNA;提取基因组DNA,PCR检测Rap1GAP第11和19号外显子(E11、E19)。SPSS 18.0软件进行统计学分析。结果 (1)正常宫颈组织中,不论是否HPV16/18感染,均能检出Rap1GAP mRNA,检出率为100%。宫颈癌组织中,HPV16/18感染标本Rap1GAP mRNA的检出率为0%,未感染标本检出率为60%,HPV16/18感染和Rap1GAP mRNA检出率呈负相关(P<0.05)。(2)正常宫颈组织和宫颈癌中E11、E19的检出率均分别为100%、90%,两组比较差异无统计学意义(P>0.05)。宫颈癌HPV16/18感染标本E11和E19的检出率均为87%,未感染标本检出率均为100%,两组比较差异无统计学意义(P>0.05)。结论 HPV16/18感染下调Rap1GAP mRNA表达可能是其导致宫颈癌的机制之一。

Objective To explore the influence of HPV16/18 infection on tumor suppressor RaplGAP in cer- vix tissue, and reveal the molecular mechanism of HPV16/18 infection causing cervical cancer. Methods 20 samples of normal cervical tissue from hysteromyoma resection and 20 samples of cervical cancer tissue were collected. After HPV16/18 was detected, the total RNA was extracted with trizol. Semi-quantitative RT- PCR was used to detect the mRNA level of RaplGAP. Genomic DNA was extracted with genomic DNA iso- lation Kit. PCR was used to detect the exons of 11 and 19 of RaplGAP. Data were analysed by SPSS 18.0 software. Results （1） The detection rate of RaplGAP mRNA was 100% in normal cervical regardless of HPV16/18 infection. In cervical cancer, the detection rate of RaplGAP mRNA was 0% in HPV16/18 infec- ted samples and 60% in non-HPV16/18 infected samples. There was a negative correlation between HPV16/18 infection and the detection rate of RaplGAP mRNA in cervical cancer （P〈0.05）. （2） The de- tection rates of exons 11 and 19 of RaplGAP were 100% and 90% respectively in both groups with no sig- nificant difference （P〉0.05）. In cervical cancer samples with HPV16/18 infection, the detection rates of both Eli and El9 were 87%, while 100% in samples without HPV16/18 infection; there was no significant difference between the two groups （P〉0.05）. Conclusion The down-regulation of RaplGAP at mRNA by HPV16/18 infection may be one of the mechanisms of cervical cancer generation.