皮肤鳞状细胞癌组织中miR-21、miR-31、Ras GTP酶激活蛋白1表达与人乳头瘤病毒的相关性研究

Association of Human papillomavirus with expression levels of miR-21, miR-31, RAS GTPase activating protein 1 in cutaneous squamous cell carcinoma

目的:测定皮肤鳞状细胞癌(CSCC)组织中人乳头瘤病毒(HPV)、miR-21及miR-31的表达量,分析HPV感染与miRNA之间的相关性,并探讨miR-31可能的致癌机制。方法:采用HPV DNA检测试剂盒和巢式聚合酶链式反应(PCR)检测CSCC组和正常对照组中HPV表达情况,实时荧光定量PCR分别测定HPV阳性和阴性CSCC组织中miR-21及miR-31的表达量,免疫组化检测CSCC组织中Ras GTP酶激活蛋白(RASA)1表达水平。结果:CSCC组织中HPV检出率为18.29%(15/82)。miR-21和miR-31在CSCC组织中相对表达量分别为0.2750±0.2280和0.0130±0.0220,均高于正常对照组(0.0110±0.0190、0.0002±0.0002),差异均有统计学意义(P均<0.05);RASA1蛋白在CSCC组织中表达明显减少(P<0.05)。在HPV阴性组织中RASA1蛋白表达与miR-31水平呈负相关(r=-0.588,P<0.05)。结论:HPV可能在CSCC发展早期起一定作用;miR-31可能是HPV相关性miRNA;高表达的miR-31可能通过下调RASA1促进细胞增殖,参与CSCC的发生及发展。

Objective： To assess the expression levels of Human papillomavirus （HPV）, miR-21 and miR-31 in cutaneous squamous cell earcinoma（CSCC） and to determine the correlation between HPV infection and miRNA in order to explore the carcinogenic mechanism of miR-31 in CSCC. Methods： HPV DNA detection kit and nested PCR were used to detect HPV in CSCC and normal controls. Real-time quantitative PCR was used to determine the expression levels of miR-21 and miR- 31 in HPV-positive and HPV-negative CSCC. The expression levels of RASA1 in CSCC was detected by immunohistochemistry. Results： The positive rate of HPV in CSCC was 18.29%（15/82）. Expression levels of both miR-21 and miR-31 were significantly higher in CSCC tissue than in normal controls（0.2750±0.2280 vs. 0.0110±0.0190 for miR-21, 0.0130±0.0220 vs. 0.0002±0.0002 for miR-31, P〈0.05 for both）. In contrast, expression levels of RASA1 were significantly lower in CSCC tissues （P〈0.05 vs. controls）. RASA1 expression levels were negatively correlated with the expression levels of miR-31 in HPV-negatire tissues（r=-0.588, P〈0.05）. Conclusions： HPV might play a role in the development of CSCC at early stage, miR-31 could be associated with HPV infection. The high expression levels of miR-31 can down-regulate RASA1 protein expression, leading to stimulation of cell proliferation and the development