喉组织中HPV11,16 E6基因DNA及HPV16 E6 mRNA的定量检测

①目的探讨人乳头瘤病毒11,16型(HPV11,16)转化基因E6 DNA及HPV16 E6 mRNA与喉疾病严重程度的关系.②方法分别用竞争性聚合酶链反应(CPCR)和半定量RT-PCR技术检测喉乳头状瘤、喉鳞癌组织中HPV11,16 E6 DNA和HPV16 E6 mRNA的含量.③结果喉鳞癌组织中HPV11,16 E6 DNA和HPV16 E6 mRNA的含量均高于喉乳头状瘤组织,差异有显著性(t=3.745,2.776,2.759, P<0.01,0.05);高、中、低分化喉鳞癌组织中HPV11,16 E6 DNA含量无显著性差异(F=0.335, 0.234, P>0.05),但低分化喉鳞癌组织中HPV16 E6 mRNA的表达水平显著高于高分化喉鳞癌(F=4.219,q=4.107, P<0.05).④结论 HPV11,16 E6 DNA和HPV16 E6 mRNA的含量有随病理程度加重而增高的趋势,从分子水平探讨转化基因E6的致癌性, 有助于阐明HPV与喉癌发生发展的关系.

DETECTION OF HPV11,16 DNA AND HPV16 E6 mRNA IN LARYNGEAL TISSUE WITH QUANTITATIVE POLYMERASE CHAIN REACTION

Objective\ To explore the relationship between disease stage and quantities of human papillomavirus type 11 and 16(HPV11 and 16) transforming gene E6 DNA and HPV16 E6 mRNA in laryngeal tissue. \ Methods\ The quantities of HPV11, 16 E6 DNA and the expression of HPV16 E6 mRNA in laryngeal papilloma and laryngeal squamous cell carcinoma tissues were detected with competitive PCR(CPCR) and semi quantitative RT PCR.\ Results\ Both the DNA copies of HPV11 and HPV16 and the expression of HPV16 E6 mRNA in laryngeal carcinomas were significantly higher than those of laryngeal papillomas, showing significant difference ( t=3.745,2.776,2.759,P <0.01, 0.05). The difference of HPV11 or HPV16 DNA copies was not significant between well differentiated squamous cell carcinoma(WDSCC), moderately differentiated squamous cell carcinoma(MDSCC) and poorly differentiated squamous cell carcinoma(PDSCC)( F=0.335, 0.234, P >0.05). However, the expression of HPV16 E6 mRNA in PDSCC was higher than that of WDSCC. \ Conclusion\ The quantities of HPV11, 16 E6 DNA and HPV16 E6 mRNA in laryngeal tissue tend to increase along with the course of laryngeal disease. It proves the carcinogenicity of transforming gene E6 at the molecular level and contributes to clarify the relationship between HPV infection and laryngeal carcinomas.