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目的:探讨Period1在肝细胞癌(HCC)组织中的表达及意义。方法:用RT-PCR与Western blot法检测30例HCC组织与其癌旁肝组织及5例正常肝组织中Period1 mRNA与蛋白表达;甲基化特异性PCR检测HCC组织与癌旁肝组织中Period1启动子甲基化水平;免疫组化检测67例HCC组织及其癌旁肝组织石蜡切片中Period1蛋白的表达;分析Period1的表达与HCC患者临床病理特征及预后的关系。结果:HCC组织Period1 mRNA和蛋白的表达水平均较癌旁肝组织与正常肝组织明显降低(均P<0.05),而两者在癌旁肝组织与正常肝组织间差异无统计学意义(均P>0.05);HCC组织Period1启动子甲基化水平较其癌旁肝组织明显增高(均P<0.05)。HCC组织石蜡切片中Period1阳性表达率为47.8%,而癌旁肝组织阳性表达率为83.6%,差异有统计学意义(P<0.05);Period1表达与HCC分化程度、结节数目、有无静脉浸润有关(均P<0.05);Period1阳性表达HCC患者中位复时间与中位生存时间均明显短于阳性者(均P<0.05)。结论:Period1在肝癌中表达下调可能与HCC的发生发展和侵袭转移密切相关;启动子甲基化可能是Period1表达下调的重要机制。
Objective: To investigate the expression and significance of Period1 in hepatocellular carcinoma (HCC). METHODS: The expression of Period1 mRNA and protein in 30 HCC tissues, adjacent non-cancerous liver tissues and 5 normal liver tissues were detected by RT-PCR and Western blot. The methylation-specific PCR was used to detect the expression of Period1 mRNA in HCC tissues and adjacent non-cancerous liver tissues The methylation level of Period1 was detected by immunohistochemistry in 67 cases of HCC tissues and para-cancerous liver tissues. The relationship between the expression of Period1 and the clinicopathological features and prognosis of HCC was analyzed. Results: The expression level of Period1 mRNA and protein in HCC tissues were significantly lower than those in paracancer tissues and normal liver tissues (all P <0.05), but there was no significant difference between the adjacent tissues and normal liver tissues P> 0.05). The methylation level of Period1 promoter in HCC tissues was significantly higher than that in adjacent non-cancerous liver tissues (all P <0.05). The positive rate of Period1 in paraffin sections of HCC tissues was 47.8%, while the positive rate of para-carcinoma in liver tissues was 83.6% (P <0.05). The expression of Period1 correlated with the degree of HCC differentiation, the number of nodules, (All P <0.05). The median recovery time and median survival time in patients with Period1-positive HCC were significantly shorter than those with positive (P <0.05). Conclusion: The down-regulation of Period1 in hepatocellular carcinoma may be closely related to the occurrence, development and invasion and metastasis of HCC. Promoter methylation may be an important mechanism of down-regulation of Period1 expression.