阿霉素对肝癌细胞系MHCC97-L中Wnt5b和Nanog表达的影响

Effects of Adriamycin on the Expression of Wnt5b and Nanog in Hepatocellular Carcinoma Cell Line MHCC97-L

目的:探讨干细胞相关基因Wnt5b和Nanog在阿霉素(ADM)作用下的人低转移性肝癌细胞系MH-CC97-L中的表达。方法:MHCC97-L细胞常规培养,用四甲基偶氮唑盐比色法(MTT)检测ADM作用72 h时MHCC97-L的半数致死量(LD50);蛋白质免疫印迹法(Western blot)检测ADM(LD50)作用下,干细胞相关基因Wnt5b和Nanog蛋白表达。结果:MTT实验结果显示,ADM对MHCC97-L的半数致死量(ID50)为0.423 3μmol/L;Western blot检测发现,在ADM(ID50)作用下,随作用时间延长,Wnt5b蛋白表达水平逐渐增高,12 h时达最高峰(P<0.05),以后则逐渐降低,72 h时降到最低(P<0.05);在ADM(ID50)作用下,随作用时间延长,Nanog蛋白表达水平逐渐降低,12 h时降到最低,以后则逐渐升高,72 h时达到最高(P<0.05)。结论:干细胞相关基因Wnt5b、Nanog在ADM作用下的人肝癌细胞系MHCC97-L中均有不同程度的表达。

Objective： To explore the expression of stem cell associated gene WntSb and Nanog in hepatocellular carcinoma cell line MHCC97-L incubating with adriamycin （ADM）. Methods： MH- CC97-L cells were routinely cultured. Median lethal dose （ LD50 ） of MHCC97-L treated with ADM for 72 h were determined with MrlT method. The expression of WntSb and Nanog proteins in MHCC97-L after treating with ADM （ LDso ） was detected with Western blot. Results： MTY results showed that LDso of MHCC97-L treated with ADM was 0. 423 3 μmol/L. The expression of WntSb increased when MHCC97-L was exposed to ADM within 4 hours, and reached peak at 12 hours, and then decreased gradually, and reduced to minimum at 72 h. When the incubation time was less than 12 h, Nanog de- creased gradually with the time passing（P 〈0. 05）, and reduced to minimum at 12 h, and then it in- creased gradually, reached peak at 72 hours. Conclusions： WntSb and Nanog proteins express in MH- CC97-L exposed to ADM in different degrees.