CAJ | 학술논문

目的了解细胞角蛋白18(CK18)磷酸化水平与HBV感染之间的关系.方法选取CHB患者肝活组织标本21份,另选肝移植健康供体肝组织14份作为对照,免疫荧光双染法和Western印迹检测其CK18磷酸化丝氨酸(pSer) 33及pSer52水平的变化;利用HepG2细胞系转染1.3 mer HBV载体或者不含HBV的空载体,免疫荧光双染法和Western印迹检测其CK18 pSer33及pSer52水平的变化,提取细胞总mRNA,实时定量PCR相对定量分析细胞分化周期因子2(cdc2),蛋白激酶Cε(pKCε)的表达水平.两独立样本间采用t检验.结果与健康对照者相比,CHB患者HBsAg染色的肝细胞,CK18 pSer33水平显著增高(t=6.618,P=0.000),而pSer52差异无统计学意义(t=2.429,P=0.051);转染HBV的HepG2细胞CK18 pSer33水平,cdc2 mRNA水平均显著高于转染空载体的细胞(t=5.365,P=0.006),而pSer52和pKCεmRNA水平则差异无统计学意义(t=1.098,P=0.334).结论 HBV感染与CK18 pSer33水平的变化密切相关.
목적 료해세포각단백18(CK18)린산화수평여HBV감염지간적관계.방법 선취CHB환자간활조직표본21빈,령선간이식건강공체간조직14빈작위대조,면역형광쌍염법화Western인적검측기CK18린산화사안산(pSer) 33급pSer52수평적변화;이용HepG2세포계전염1.3 mer HBV재체혹자불함HBV적공재체,면역형광쌍염법화Western인적검측기CK18 pSer33급pSer52수평적변화,제취세포총mRNA,실시정량PCR상대정량분석세포분화주기인자2(cdc2),단백격매Cε(pKCε)적표체수평.량독립양본간채용t검험.결과 여건강대조자상비,CHB환자HBsAg염색적간세포,CK18 pSer33수평현저증고(t=6.618,P=0.000),이pSer52차이무통계학의의(t=2.429,P=0.051);전염HBV적HepG2세포CK18 pSer33수평,cdc2 mRNA수평균현저고우전염공재체적세포(t=5.365,P=0.006),이pSer52화pKCεmRNA수평칙차이무통계학의의(t=1.098,P=0.334).결론 HBV감염여CK18 pSer33수평적변화밀절상관.
Objective To investigate the relationship between hepatitis B virus (HBV) infection and cytokeratin 18 (CK18) phosphorylation.Methods Liver tissues were taken from 21 chronic hepatitis B (CHB) patients by liver biopsy and 14 healthy controls were collected.Immunofluorescence double staining and Western blotting were used to detect CK18 pSer33 and CK18 pSer52 phosphorylation.HepG2 cell line was transfected with either 1.3 mer HBV vector or empty control vector.CK18 pSer33 or CK18 pSer52 phosphorylation were tested using immunofluorescence double staining and Western blotting.Total mRNA was extracted from the cells.The expressions of cell division cycle 2 (cdc2) and protein kinase Cε (pKCε) were analyzed by realtime polymerase chain reaction relative quantification.Statistical analyzee were performed by using two-independent samples t test.Results Compared to healthy controls,phosphorylation of CK18 pSer33 in HBsAg stained hepatocytes was significantly higher in CHB patients (t=6.618,P=0.000).However,no difference was found in phosphorylation of pSer52 (t=2.429,P=0.051).Phosphorylation of CK18 pSer33 and expression of cdc2 mRNA in HBV transfected HepG2 were significantly higher in empty vector transfected HepG2 (t=5.365,P=0.006),while no difference was found in phosphorylation of pSer52 and expression of pKCε mRNA (t=1.098,P=0.334).Conclusion HBV infection is significantly associated with phosphorylation of CK18 pSer33.