小剂量5-脱氧杂氮胞苷诱导HepG2肝癌细胞新生肿瘤-睾丸抗原CT10和SSX1表达及其意义

Expression and significance of novel tumour-testis antigens CT10,SSX1 induced by low dose of 5-aza-2’-deoxycytidine in hepatocellular carcinoma cell HepG2

目的探讨小剂量5-脱氧杂氮胞苷(5-aza-CdR)诱导HepG2肝癌细胞肿瘤-睾丸抗原(nCTAs)的CT10和SSX1表达及其意义。方法采用RT-PCR方法检测HepG2、SMMC-7721、BEL-7402、MHCC97L和MHCC97M3肝癌细胞株MAGE1、MAGE3、CT10、CTp11、NE-ESO-1和SSX16种CTAs的mRNA表达,挑选不表达CT10和SSX1的HepG2细胞作为nCTAs表达的研究对象;用0、0.5、1.0、1.5μmol/L 5-aza-CdR处理HepG2细胞,不含5-aza-CdR为对照组,从低到高浓度分别为实验1、2、3组;分别采用RT-PCR和Western blot检测CT10和SSX1 mRNA和蛋白表达。结果5株肝癌细胞均存在不同的nCTAs表达,表达呈异质性改变,其中HepG2表达MAGE1、MAGE3、CTp11和NY-NSO-1,MHCC97L和MHCC97M3表达MAGE1、MAGE3、CT10、CTp11和SSX1,BEL-7402表达MAGE1、MAGE3和CTp11,而SMMC-7721只表达MAGE1、MAGE3。实验1、2、3组均能诱导新生CTA-CT10和SSX1表达,其中0.5μmol/L小剂量的实验1组也能诱导nCTAs的表达;实验3组诱导的nCTAs相对表达量显著高于实验1、2组,差异有统计学意义(P<0.05)。结论不同的肝癌细胞表达不同的nCTAs,小剂量5-aza-CdR能诱导肝癌细胞株表达nCTAs。

Objective To explore expression and significance of novel tumour-testis antigens CT10,SSX1 induced by low dose of 5-aza-2’-deoxycytidine(5-aza-CdR)in hepatocellular carcinoma cell HepG2.Methods The six mRNA expression of MAGE1,MAGE3,CT10,CTp11,NE-ESO-1,and SSX1 in HepG2,SMMC-7721,BEL-7402,MHCC97L and MHCC97M3 liver cancer cell lines were detected by RT-PCR.HepG2 cells that do not express CT10 and SSX1 were selected as the study object as nCTAs,and HepG2 cells were treated with 0,0.5,1.0,and 1.5μmol/L 5-aza-CdR.The control group did not contain 5-aza-CdR,from low to high concentration,they were experiment 1,2,and 3 groups respectively,the mRNA and protein expressions of CT10 and SSX1 were detected by RT-PCR and Western blot,respectively.Results Different nCTAs expression was found in five hepatocellular cells,the expression was heterogeneous,among them MAGE1,MAGE3,CTp11,and NY-ESO-1 expression in HepG2;MAGE1,MAGE3,CT10,CTp11,and SSX1 expression in both MHCC97L and MHCC97M3;MAGE1,MAGE3,and CTp11 expression in BEL-7402;however,SMMC-7721 only expresses MAGE1 and MAGE3.Experimental 1,2,and 3 groups could induce the expression of CTA-CT10 and SSX1,among them,0.5μmol/L small dose of experiment group 1 could also induce the expression of nCTAs,the relative expression of nCTAs induced in experiment 3 group was significantly higher than that in experiment 1 and 2 group,and the differences were statistically significant(P<0.05).Conclusion Different hepatocellular carcinoma cells expressed different CTAs,and a small dose of 5-aza-CdR could induce the expression of nCTAs in hepatocellular carcinoma cell lines.