AFP、GPC3、ZHX2及ZBTB20基因在小鼠肝脏再生过程中的表达及其意义

Expression and significance of AFP,GPC3,ZHX2,ZBTB20 in liver regeneration of mice

目的探讨小鼠肝脏再生过程中甲胎蛋白(AFP)、磷脂酰肌醇蛋白聚糖￣3(GPC3)、锌指和同源框2(ZHX2)、锌指蛋白ZBTB20基因的表达及其意义。方法 30只C57/BL6小鼠,按随机数字表法分成肝脏再生模型组(肝再生组)和肝微量切除组(肝微切组),各15只。肝再生组切除肝组织约占肝脏质量的70%;肝微切组切除肝组织约占肝脏质量的5%。分别取两组小鼠肝切除时(0 h)及切除术后2、4、24、48及72 h的肝脏组织,应用实时荧光定量聚合酶链反应(RQ-PCR),检测不同时间点甲胎蛋白(AFP)信使核糖核酸(mRNA)表达;同时应用RQ-PCR法检测24、48及72 h的肝脏组织GPC3、ZHX2、ZBTB20 mRNA表达。采用t检验比较各组AFP、GPC3、ZHX2、ZBTB20 mRNA表达变化。结果肝再生组AFP mRNA术后2、4、24、48及72 h分别为0 h正常肝组织基因表达的(70±20)%、(80±20)%、(90±10)%、(240±40)%、(870±120)%。72 h的AFP基因表达变化与24、48 h相比较,差异均有统计学意义(t=11.2、8.6,P=0.001、0.001),术后24 h AFP表达开始上升;术后24、48、72 h的GPC3mRNA分别为0 h正常肝组织基因表达的(50±10)%、(120±20)%、(190±50)%。72 h的GPC3基因表达变化与24 h相比较,差异有统计学意义(t=4.8,P=0.01),术后48 h GPC3表达开始上升;ZHX2、ZBTB20mRNA在术后24、48 h分别为0 h正常肝组织基因表达的(50±10)%,(60±10)%和(70±20)%,(50±10)%。72 h的ZHX2升高(120±30)%,与24、48 h相比,差异有统计学意义(t=3.8、3.3,P=0.04、0.04)。72 h的ZBTB20升高(140±30)%,与24、48 h相比,差异有统计学意义(t=3.4、4.9,P=0.04、0.01)。结论小鼠肝大部切除术后肝脏再生启动,AFP、GPC3基因可能在肝脏再生中参与了细胞增殖的主动调控,转录抑制因子ZHX2、ZBTB20 mRNA在肝脏再生中表达下调,对AFP、GPC3启动子的转录抑制作用下降,从而促进肝组织再生,对肝脏再生具有一定的意义。

Objective To study the expression and significance of alpha-fetoprotein （AFP）, glypican-3 （GPC3）, zinc fingers and homeoboxes （ZHX） 2 and zinc finger protein ZBTB20 in mouse liver regeneration. Methods Thirty C57/BL6 mice were divided into liver regeneration model group and liver trace resection group with 15 mice per group. The mice in the liver regeneration model group underwent 70% liver resection and the mice in the liver trace resection group underwent 5% liver section. The liver tissues fight after resection （0 h） and 2, 4, 24, 48, 72 h after resection were obtained and the expression of AFP messenger ribonucleie acid （mRNA） were tested by real time reverse transcription polymerase chain reaction analysis. The expression of GPC3, ZHX2, ZBTB20 mRNA of liver tissues at 24, 48, 72 h after resection were also tested by real time RT-PCR analysis and the difference between 2 groups were compared by t-test. Results The AFP mRNA expression levels at 2, 4, 24, 48, 72 h after liver resection were （70±20）%, （80±20）%, （90±10）%, （240±40）%, （870±120）% respectively compared with the level of the normal liver tissues. There was significant difference between the AFP gene expression of liver tissues at 72 h after resection and 24, 48 h after resection （t=11.2,8.6, P=0.001, 0.001）. The GPC3 mRNA expression 24, 48, 72 h after liver resection were （50±10）%, （120±20）%, （190±50）% of the normal liver tissues. The AFP and GPC3 expression increased obviously at 24 h and 48 h after resection. There was significant difference between GPC3 gene expression in the liver tissues of 72 h and 24 h after resection （t=4.8, P=0.01）. ZHX2, ZBTB20 mRNA expression in the liver tissues of 24 h and 48 h were （50±10）%, （60±10）% and （70±20）%, （50：t：10）% of the normal liver tissues. The ZHX2 expression in the liver tissues of 72 h rose by （120±30）% and there was significant difference compared with that of 24 h, 48 h （t=3.8,3.3, P=0.04,0.04）. The ZBTB20 expression in the liver tissues of 72 h rose by （140±30）% and there was significant difference compared with that of 24 h, 48 h （t=3.4,4.9, P=0.04,0.01）. Conclusions The liver regeneration is activated after partial hepatectomy. AFP and GPC3 may play a role in the initiative regulation of cell proliferation during the liver regeneration. The expression of ZHX2, ZBTB20 mRNA are down-regulated during the initial state and thus the transcription inhibition of AFP and GPC3 decline so thus promote the liver regeneration.