论文部分内容阅读
目的检测肝细胞癌患者肝组织中乙型肝炎病毒共价闭合环状DNA(HBV ccc DNA),探讨其在肝细胞癌发生、发展中的意义。方法选取20例肝细胞癌患者的癌及癌旁组织,经10%福尔马林固定、石蜡包埋、连续切片,采用0.05%多聚赖氨酸进行处理。经不降解质粒的ATP依赖的DNA酶(PSAD酶)消化后,应用原位滚环扩增(insitu RCA)结合原位跨缺口PCR扩增技术检测肝组织中ccc DNA的表达。结果 20例肝癌患者标本中有17例检测到HBV ccc DNA阳性信号,为蓝色或紫蓝色并呈团块状,定位于细胞核。肝癌患者癌及癌旁组织HBV ccc DNA阳性率分别为10%(2/20)和85%(17/20),差异具有显著统计学意义(P<0.01)。结论肝细胞癌患者癌旁组织病毒复制水平高于癌组织。
Objective To detect the hepatitis B virus covalently linked circular DNA (HBV ccc DNA) in hepatocellular carcinoma (HCC) patients and to explore its significance in the occurrence and development of hepatocellular carcinoma. Methods Twenty cancer and paracancerous tissues from patients with hepatocellular carcinoma were selected and fixed in 10% formalin and embedded in paraffin. The cells were serially sectioned and treated with 0.05% polylysine. After enzymatic digestion with ATP-dependent DNase (PSADase), which did not degrade plasmids, the expression of ccc DNA in liver tissue was detected by in situ rolling-in amplification (insitu RCA) and in situ cross-notch PCR amplification. Results The positive signal of HBV ccc DNA was detected in 17 of the 20 HCC patients, which were blue or purple-blue and clustered in the nucleus. The positive rates of HBV ccc DNA in cancer tissues and adjacent tissues were 10% (2/20) and 85% (17/20), respectively, with significant difference (P <0.01). Conclusion Hepatocellular carcinoma patients with paracancerous tissue virus replication level is higher than the cancer tissue.