PCR和原杂交技术检测鼻咽癌石蜡组织中HHV-6DNA

Detection of Human Herpesvirus 6 in Paraffin-Embeded NPC Tissues by PCR and ISH

为了解人疱疹病毒６型（ＨＨＶ－６）与鼻咽癌（ＮＰＣ）的可能关系，检测了ＮＰＣ发生不同阶段的石蜡组织中ＨＨＶ－６ＤＮＡ的存在情况。方法：采用ＰＣＲ、斑点杂交和原位杂交技术，共检测了４２例鼻咽癌石蜡组织、２１例鼻咽癌前病变石蜡组织和２７例正常鼻咽石蜡组织ＨＨＶ－６ＤＮＡ的存在情况。结果：通过ＰＣＲ扩增，在４２例ＮＰＣ组织中检出１３例存在ＨＨＶ－６ＤＮＡ，占３１％；２１例鼻咽癌前病变组织中１例阳性（４７％），而２７例正常鼻咽组织中无一例阳性。斑点杂交的结果表明ＰＣＲ扩增是ＨＨＶ－６特异的。原位杂交发现，在１３例ＰＣＲＨＨＶ－６阳性的ＮＰＣ组织中１１例ＨＨＶ－６ＤＮＡ阳性。ＨＨＶ－６ＤＮＡ主要分布于癌巢的肿瘤细胞核内，少数亦可见于癌旁的异型细胞，但在淋巴细胞及正常上皮细胞内未发现阳性信号。结论：本实验用分子生物学技术首次证明，在中国ＮＰＣ患者的鼻咽石蜡组织中存在ＨＨＶ－６。ＨＨＶ－６与ＮＰ相关。

Objective:In attempt to understand the possible relationship between HHV-6 and NPC,we investigate the presence and distribution of HHV-6 in tissues of NPC. Methods: PCR and dot blot hybridiation and in situ Hybridization (ISH) were applied to detect the presence and distribution of HHV-6 DNA in 42 tissues of NPC,21 of precancerous nasopharynx and 27 of normal nasopharynx.Results: HHV-6 DNA were detected in 31% (13/42) tissues of NPC,4 7%(1/21) of precancerous nasopharynx and none of normal nasopharynx.The result of dot blot hybridization confirmed the HHV-6 specificity of PCR.By ISH,HHV-6 were only detected in 11 tissues of NPC.In addition,ISH showed that positive HHV-6 were mainly restricted to neoplastic cells.In no case the HVV-6 positive was detected in lymphocyte and normal epithelial cells.Conlusion: This study provides molecular evidence of presence and distribution of HHV-6 in tssues of NPC patients in China for the first time . HHV-6 is associated with NPC and appears to play a special role in the pathogenesis of NPC.