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目的 采用多种方法检测丙型肝炎病毒 (HCV)核心区基因和表达产物在肝细胞肝癌(HCC)组织中的分布及其在HCC发生和发展中的作用。方法 对 39例HCC患者进行免疫组化和原位杂交的检测。IS RT PCR法用于检测并定位HCVRNA。微切组织的RT PCR法分别检测癌与癌旁组织中的HCVRNA ,以便能够控制扩增产物的来源。同时还进行血清学ELISA和RT PCR的检测。结果 血清ELISA的阳性率 (30 8% )并不总与RT PCR的阳性率 (43 6 % )一致 ,后者对于反映HCRRNA存在更为敏感和准确。免疫组化结果显示HCV核心抗原的表达率为 38 5 %。阳性信号位于癌旁肝细胞的胞浆 ,而在癌组织中发生了显著的核转位表达 (73 3% )。IS RT PCR结果显示阳性信号位于肝 (癌 )细胞的胞浆。阳性率 (5 3 8% )与血清中的HCVRNA的水平显著相关。但有 4例肝内HCVRNA高表达而血清HCVRNA的水平很低或是阴性 ,提示血清中HCVRNA的水平并不总能反映肝内HCVRNA的水平。微切组织RT PCR法的检测结果显示癌与癌旁组织中HCVRNA的阳性率一致 ,为 5 9 0 %。结论 HCVRNA在HCC组织中的高表达提示HCVRNA在肝细胞恶性转化中起着重要的作用。IS RT PCR是一个理想的检测并定位HCVRNA的方法。微切组织的RT PCR具有与众不同的优点 ,我们期待这种方法为确定HCV在肝细胞恶性转?
Objective To detect the distribution of hepatitis C virus (HCV) core region genes and their expression products in hepatocellular carcinoma (HCC) tissues by multiple methods and their roles in the development and progression of HCC. Methods 39 cases of HCC patients were detected by immunohistochemistry and in situ hybridization. The IS RT PCR method is used to detect and localize HCV RNA. The microtissue RT PCR method detects the HCVRNA in the cancer and paracancerous tissues, respectively, in order to be able to control the source of the amplification product. Serological ELISA and RT PCR were also performed. Results The positive rate of serum ELISA (30.8%) was not always consistent with the positive rate of RT PCR (43.6%), which was more sensitive and accurate for reflecting the presence of HCRRNA. The results of immunohistochemistry showed that the expression rate of HCV core antigen was 38.5%. Positive signals were located in the cytoplasm of paracancer hepatocytes, whereas significant nuclear translocation expression was found in cancerous tissues (73.3%). IS RT PCR results showed that the positive signal was located in the cytoplasm of liver (cancer) cells. The positive rate (53.8%) was significantly correlated with the level of HCV RNA in serum. However, in 4 cases, intrahepatic HCVRNA was highly expressed and serum HCVRNA level was low or negative, suggesting that serum HCVRNA level does not always reflect intrahepatic HCVRNA level. The results of micro-sectioned RT-PCR showed that the positive rate of HCVRNA in cancer and adjacent tissues was 59.0%. Conclusion The high expression of HCVRNA in HCC tissues suggests that HCVRNA plays an important role in the malignant transformation of hepatocytes. IS RT PCR is an ideal method to detect and localize HCVRNA. RT PCR of micro-tissue tissue has its own unique advantages. We expect this method to determine the malignant transformation of HCV in hepatocytes.