尖锐湿疣组织中微卫星DNA的改变与HPV感染的相关性

Relationship between Microsatellite Alterations in Tissues of Condyloma Acuminatum and HPV Infection

目的探讨特定位点的微卫星不稳定性(MSI)及杂合性丢失(LOH)在尖锐湿疣(CA)组织中的检出情况及其与人乳头瘤病毒(HPV)感染的关系。方法选取D3S1234,D3S1611,D9S171,D10S1687和D17S16785个微卫星位点,应用聚合酶链反应(PCR)和变性聚丙烯酰胺凝胶电泳-银染色技术对40例CA组织及11例CA癌变组织进行微卫星分析,并采用核酸分子快速导流杂交基因芯片技术检测HPV的基因分型。结果 CA及CA癌变组织中LOH的总检出率分别为22.50%(9/40)和63.64%(7/11),差异有统计学意义(P<0.05),两组中LOH发生频率最高的位点均为D9S171,其次为D17S1678位点。MSI的发生率较低,CA组中频率为10.00%(4/40),CA癌变组为18.18%(2/11)。CA组及CA癌变组标本,高危型HPV感染者LOH阳性率明显高于低危型HPV感染者,差异有统计学意义(P=0.019);两组中MSI阳性的6例样本均为高危型HPV感染。结论高频LOH位点D9S171和D17S1678连锁的抑癌基因p16,p53可能在CA癌变的发生过程中起着重要作用;另外,该结果也说明HPV感染,尤其高危基因型病毒是CA癌变的因素之一。微卫星DNA的改变与高危型HPV的感染有一定的相关性。

Objective To explore the relationship between microsatellite alterations in tissues of condyloma acuminatum（CA） and human papillomavirus （HPV） infection. Methods Five sites of microsatellite polymorphism of human gene were selected, and both microsatellite instability （MSI） and loss of heterozygosity （LOH） were tested in 40 samples of CA and 11 samples of CA with canceration by PCR-denaturing PAGEsilver staining technique. The infection genotype of HPV was also detected by flow-through rapid hybridization technique. Results The total incidence of LOH in CA and CA with canceration were 22.50% （9/ 40） and 63.64% （7/11） respectively. There was a significant difference between the two groups （P 〈 0.05）. D9S171 showed the highest incidence of LOH on the five markers, the second was D17S1678. MSI was not frequent. The MSI rate of CA and CA with canceration were 10.00% （4/40） and 18.18% （2/11 ） respectively. Based on analysis of the two groups, the rate of LOH for high-risk HPV were higher than those for low-risk HPV. There was statistical significance （P = 0. 019）. All 6 subjects which were categorized as positive MSI were also infected with high risk HPV type. Conclusion The frequencies of LOH at the microsatellite locus D9S171 in p16 gene and D17S1678 in P53 gene may be play an important role in the development of carcinogenesis of CA. In addition, HPV, particularly high-risk genotype virus, may be one of the multiple carcinogenic factors in CA. Microsatellite alterations is associated with high-risk HPV type.