[Detection of hepatitis C virus RNA in cells with hepatocellular carcinoma and its practical significance by in situ polymerase chain reaction].

Persistent infection with hepatitis C virus (HCV) is associated with chronic hepatitis and cirrhosis which may eventually develop into hepatocellular carcinoma (HCC). As far, the pathogenesis mechanism of HCC is unclear and nothing is known of the distribution, frequency or type of infected cells in HCC. One-step reverse transcription in situ polymerase chain reaction (ORT-PCRIS) to detect HCV RNA in HCC samples was developed in our laboratory. Liver tissues from 27 patients with HCC were investigated by this technique for frozen or paraffin-embedded sections. Meanwhile, HCV RNA in sera and extracts of specimens with HCC were assayed by RT-PCR. The positive rate of HCV RNA by ORT-PCRIS was 81.5% (22/27) in the peritumor liver tissues and 77.8% (21/27) in the tumor tissues, significantly higher than 29.6% (8/27) in sera and 37.0% (10/27) in the extreats of HCC by RP-PCR (P < 0.01). HCV RNA positive signals were found mainly in the nuclei of tumor cell, and mainly both the nuclei and cytoplasms in peritumor cells and mainly both the nuclei and cytoplasms in peritumor cells (P < 0.05). Positive granules of HCV RNA were much more in peritumor cells than in the tumor cells. Positive cells were scattered mainly on the point-type in the cancer tissues and on the diffusion-type in the peritumor tissues. Our findings suggest that ORT-PCRIS for detecting HCV RNA in the cell with HCC remarkably prior to traditional RT-PCR. HCV infection plays a relatively important role in determination of HCC development and perhaps HCV replication and its genomic RNA integration with hepatocyte DNA are involved in the course of the malignant transformation of hepatocytes.