ObjectiveTo observe the expression of cathepsin S(Cat-S)in hepatoma carcinoma cell and normal hepatic cell,so as to investigate the relationship between expression of Cat-S and occurrence and development of hepatocellular carcinoma(HCC).MethodsHuman HCC with high metastasis potential MHCC97-H,with low metastasis potential MHCC97-L and normal hepatic cell LO2 were cultured in vitro,then the Cat-S mRNA expression level were detected by Fluorecence quantitative PCR.ResultsCat-S mRNA was at low level in cell lines LO2,but significantly higher in cell lines MHCC97-H and cell lines MHCC97-L(P0.05);and which in in cell lines MHCC97-H was significantly higher than that in cell lines MHCC97-L(P0.05).ConclusionsCat-S expression in HCC cells with high or low metastasis potential and normal hepatic cell are different,which maybe correlated with occurrence and development of HCC;Cat-S maybe used as a biologic maker for HCC diagnosis and treatment.

目的研究高转移肝癌MHCC97细胞中Twist基因表达与Wnt信号通路的关系。方法取对数生长期MHCC97细胞分为五组,A、B组分别加入0.05 mol/L氯化锂分别培养12、24 h,C、D组予0.1 mol/L氯化锂分别培养12、24 h激活Wnt通路,E组不干预。采用RT-PCR和荧光定量PCR方法观察各组Twist mRNA表达情况。结果两种检测方法均显示Twist mRNA在A、B、C、D组表达均明显高于E组,P<0.05;A、B组,C、D组,A、C组,B、D组间比较,P均>0.05。结论MHCC97细胞中Wnt通路激活后Twist基因表达明显升高未激活,即Twist基因表达与Wnt信号传导通路激活关系密切,Wnt传导通路激活与氯化锂浓度及作用时间无明显相关性。

Objective To investigate the relationship of Twist mRNA and WNT signaling pathway in liver cancer cells.Methods Human hepatoma cell line SMMC7721 cells were cultured with with 0.05 mol/L chloride lithium for 2 h,then cultured separately for 6,12,24 h,respectively,for A1,A2,A3 groups;human hepatoma cell line SMMC7721 cells were cultured with 0.1 mol/L chloride lithium for 2 h,then cultured separately for 6,12,24 h,respectively,for B1,B2,B3 Group;In addition,a blank control group was set up.The expression of Twist mRNA in SMMC7721 was detected respectively when the WNT pathway was activated and not activated by RT-PCR and real time PCR.Result The value of Twist mRNA measured by RT-PCR method in A1,A2,A3 group were 0.38±0.05、0.60±0.08、0.69±0.06,B1,B2,B3 group were 0.56±0.05、0.59±0.06、0.69±0.04,compared with the control group(0.18±0.03),P0.05.The value of Twist mRNA measured by real time PCR method in A1,A2,A3 group were 0.43±0.01、0.56±0.01、0.67±0.01,B1、B2、B3 were 0.52±0.01、0.58±0.00、0.65±0.00,compared with the control group(0.21±0.00),P0.05.Conclusion The expression of Twist gene in liver cancer may be related to WNT pathway.