喉癌和喉乳头状瘤组织中人乳头状瘤病毒和p16蛋白的检测

Human papilloma virus infection and expression of p16 protein in laryngeal papilloma and laryngeal carcinoma

目的 探讨人类乳头状瘤病毒 (humanpapillomavirus ,HPV)感染和抑癌基因p16的失活与喉癌和喉乳头状瘤 (laryngealpapilloma ,LP)发生的相关性 ,以进一步阐明喉癌和LP的病因和发病机理。方法 收集LP 46例 ,其中成人型喉乳头状瘤 (adult onsetLP ,ALP) 2 1例 ,青少年型喉乳头状瘤(juvenile onsetLP ,JLP) 2 5例、喉癌 2 6例、癌旁正常组织 6例、声带小结 15例 ,用标记的HPV1,6,8,11,13 ,16,18,30 ,31,32 ,33 ,4 5,51通用引物直接法原位聚合酶链反应 (polymerasechainreaction ,PCR)方法和免疫组化 (SP法 )方法分别检测HPV DNA和p16蛋白。结果 ①HPV阳性率JLP组 (84% ,2 1/2 5 )显著高于ALP组 (38.1% ,8/2 1)、喉癌组 (19.2 % ,5 /2 6 )、声带小结组 (0 /15 )和癌旁组织组 (0 /6 ) (χ2 检验 ,P <0 .0 5 ) ;而ALP组、喉癌组、声带小结组和癌旁组织组之间HPV阳性率差异无显著性 (χ2 检验或Fisher精确概率法 ,P >0 .0 5 ) ;②p16蛋白的阳性率ALP组 (5 7.1% ,12 /2 1)和喉癌组 (38.5 % ,10 /2 6 )均显著低于JLP组 (88% ,2 2 /2 5 )、癌旁组织组 (10 0 % ,6 /6 )和声带小结组 (93.3 % ,14/15 ) (χ2 检验 ,P <0 .0 5 ) ;ALP组和喉癌组之间及JLP组和声带小结组之间p16蛋白阳性率无显著差别 (χ2 检验 ,P >

Objective To evaluate the role of human papilloma virus (HPV) infection and inactivation of p16 gene in laryngeal papilloma (LP) and laryngeal squamous cell carcinoma (LC) . Methods HPV consensus primers direct in situ polymerase chain reaction (ISPCR) and immunohistochemical method were applied to detect the presence of HPV genomes (1,6,8,11,13,16,18,30,31,32,33,45,51) and the expression of p16 protein respectively in 93 cases of formalin fixed, paraffin imbedded specimens, which contained 46 cases of LPs [adult onset laryngeal papilloma (ALP) 21, juvenile onset laryngeal papilloma (JLP)25], 26 cases of LCs, 6 cases of normal tissues adjacent to carcinoma, and 15 cases of vocal noduli. Results (1)The difference of positive rates of HPV DNA in JLP group (84%,21/25)and other groups were statistically significant (χ 2 test, P <0.05). The difference of positive rates of HPV DNA in ALPs(38.1%,8/21), in LCs(19.2%,5/26), in vocal noduli(0%,0/15), and in normal tissues adjacent to carcinoma(0%,0/6) were not significant statistically (χ 2 test or Fisher′s exact probability test, P >0.05). (2)The positive rates of expression of p16 protein in ALP group(57.1%,12/21)and LC group(38.5%,10/26) were significantly lower than that in vocal noduli group(93.3%,14/15), in JLP group(88%,22/25), and in normal tissues adjacent to carcinoma group (100%,6/6)(χ 2 test or Fisher's exact probability test, P >0.05). There were no significant differences of positive rates of expression of p16 protein between ALP group and LC group, and between JLP group and vocal nodule group (χ 2 test, P >0.05). (3) In LPs,the difference of positive rates of p16 protein expression between HPV positive cases and HPV negative cases was significant statistically (χ 2 test, P <0.05). In LCs , there was no difference in p16 protein expression rate between the two teams(Fisher exact probability test, P >0.05). Conclusion The pathogenesis of JLP is closely associated with HPV infection and not associated with the inactivation of p16 gene. Conversely, the pathogenesis of ALP and LC is associated with the inactivation of p16 gene and not associated with the HPV infection.