原位聚合酶链反应技术及其在消化道肿瘤相关病毒基因检测中的应用

DEVELOPMENT OF DIRECT IN SITU PCR AND MEASUREMENT OF TUMOR-ASSOCIATED - VIRUS GENES IN GASTROINTESTINAL CANCER TISSUES

为克服核酸原位杂交技术(ISH)和聚合酶链反应(PCR)的不足,我们应用分子病理学原理建立了原位PCR(PCRIS)、反转录原位PCR(RT—PCRIS)技术,对27例原发性肝癌(HCC)组织中丙型肝炎病毒核酸(HCV RNA)、54例胃癌(GC)组织中EPstein—Barr病毒脱氧核糖核酸(EBV DNA)及20例食道癌(EC)组织中人乳头状瘤病毒DNA(HPV DNA)进行原位分析。结果显示HCV RNA、EBVDNA及HPV DNA阳性率分别为78％、29％及25％,明显高于ISH检测结果(P<0.05)。提示本研究建立的PCRIS、RT—PCRIS技术特异、灵敏、定性、定位;HCV、EBV、HPV感染分别与HCC、GC及EC发生、发展密切相关。

In order to go further Into the relationship between tumor -associated -virus (TAV) infection and gastrointestinal cancer(GIC), our laboratory developed a kind of polymerase chain reaction in situ(PCRIS) and reverse transcription in situ PCR(RT - PCRIS) techniaue. The techniaue was employed in studying the cellular distribution of TAV genes (HCV RNA,HPV DNA, EBV DNA) in GIC tissues, including 27 patients with hepatocellular carcinoma (HCC) , 54 gastric carcinoma (GC) and 20 esophagal carcinoma (EC). The results showed that our methods decreased the reactive times,increased the detective sensitivity and archieved the purposes of precise localization and satisfactory preservation of the cell and tissue structure. The detected rates of HCV RNA in HCC, EBV DNA in GC and HPV DNA in EC tissues by RT-PCRIS or PCRIS were 78% ,29% and 25% ,respectively, significantly higher than those by ISH. Our results suggest that RT -PCRIS or PCRIS provide an useful technique for the profound study on the carcinogenesis of TAV and confirm that HCV, EBV and HPV infection are related to HCC,GC and EC,respectively.