摘要
目的通过检测PCNA、p53及和CyclinB1 mRNA在兔肝内胆管结石胆管组织中的表达,探讨肝内胆管结石发生的分子机制。方法建立肝内胆管结石动物模型,采用半定量逆转录聚合酶链反应(RT-PCR)的方法检测30例胆管结石兔和20例正常兔胆管组织CyclinB1 mRNA的表达水平,采用免疫组织化学法检测胆管组织PCNA和p53的表达,应用流式细胞仪测定细胞周期和细胞凋亡情况。结果胆管结石兔胆管组织PCNA表达阳性率40%,明显高于对照组0,两者相比差异有统计学意义(P<0.01),胆管结石兔胆管组织p53表达阳性率0,对照组p53表达阳性率0,两者相比差异无统计学意义(P>0.05),胆管结石兔胆管组织CyclinB1 mRNA表达水平(0.436±0.133)与正常对照组(0.178±0.085)比较差异有统计学意义(P<0.01),胆管结石兔胆管G1/S期细胞比例(50.11±6.34)%,与正常组(75.23±5.29)%比较差异有统计学意义(P<0.05),G2/M期细胞比例(35.59±4.68)%,与正常组(15.35±3.76)%比较差异有统计学意义(P<0.05),胆管细胞增殖指数(18.87±1.66),与正常组(5.32±0.41)比较差异有统计学意义(P<0.05),细胞凋亡率(15.24±0.87)%,与正常组(2.33±0.36)%比较差异有统计学意义(P<0.05)。结论PCNA、p53及CyclinB1mRNA表达与肝内胆管结石生物学行为密切相关,胆管细胞大量增殖在肝内胆管结石形成中发挥重要作用,但细胞增殖与恶性细胞的无限增殖有根本的区别。
Objective To investigate the expression of PCNA, p53 and CyclinB1 mRNA in intrahepatic duct tissue in rabbit hepatolithiasis and molecular mechanism of hepatolithiasis. Methods Intrahepatic duct stone model was established in rabbits. The expression of CyclinB1 mRNA was detected by reverse transcription PCR in 30 cases of hepatolithiasis and 20 cases of normal tissue and that of PCNA and p53 by immunohistoehemistrical method. Flow cytometry was used to detect cell cycle and apoptosis. Results The expression of PCNA （40 % ） was significantly higher in experimental group than that （0） in controls （P 〈 0.01 ）. But there was no obvious significant difference in the expression of p53 between the experimental group and the control group （P 〉0.05）. The expression of CyclinB1 mRNA （0. 436±0.133） was significantly higher in experimental group than that （0. 178±0.085） in controls （P〈 0.01）. The percentage of G1/S phase （50.11 ±6.34） % was decreased remarkably in experimental group as compared with control group （75.23 ± 5.29） %, while the percentage of G2/M phase （35.59±4.68）% was increased remarkably in experimental group as compared with control group （15.35±3.76） %. The proliferative index （18.87± 1.66） in experimental group was significantly higher than that （5.32±0.41） in control group. In addition, the apoptotic rate in experimental and control groups was （ 15.24±0.87） % and （2.33 ±0.36） % respectively, with the difference being significant between them （P〈0.05）.Conclusion There is a close relationship between the expression of PCNA, p53 and CyclinB1 mRNA in intrahepatic duct tissue and the biological behavior of hepatolithiasis. The abnormal proliferation of biliary epithelium cellsplays an important role in the formation of hepatolithiasis.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第11期1296-1298,共3页
Chinese Journal of Experimental Surgery
