摘要
目的比较肝癌耐药株HepG2/ADM与其亲代HepG2的TRAIL受体的表达。及TRAIL对肝癌耐药株HepG2/ADM的治疗作用。方法通过浓度递增法用阿霉素处理人肝癌细胞株HepG2获得肝癌耐药株HepG2/ADM。RT-PCR、Western Blot和免疫组化比较肝癌耐药株HepG2/ADM与其亲代HepG2的TRAIL受体及bcl-2、MDR的mRNA和蛋白表达。采用AnnexinV-FITC-PI双染色流式细胞仪测TRAIL对肝癌耐药株HepG2/ADM的调亡诱导作用。结果肝癌耐药株HepG2/ADM与其亲代HepG2比较都存在DR4、DR5受体且mRNA和蛋白表达上无差异。DcR1、DcR2在蛋白表达上无差异,但mRNA水平有轻度变化,未达到统计显著性。免疫组化示肝癌耐药株HepG2/ADM与其亲代HepG2都存在DR4、DR5受体和DcR1、DcR2受体,分析未见差异。TRAIL对HepG2/ADM及其亲代均有诱导调亡的作用,同样存在一定的耐受现象。但调亡作用两者无显著性差异。结论肝癌耐药株HepG2/ADM与其亲代HepG2都存在DR4、DR5受体和DcR1、DcR2受体。表达无显著性差异。单纯TRAIL治疗对耐药株作用与亲代细胞作用相仿,虽同样存在一定的耐受现象,但受HepG2/ADM耐药性的影响小。TRAIL治疗肝癌耐药株有优势。
Objective To compare the expression of TRAILR in HepG2/ADM with that of HepG2 and then to investigate the therapeutic effect of TNF-related apoptosis-inducing ligand （TRAIL） in hepatocelluar carcinoma cell line HepG2/ADM which was establised to resist to chemotherapy. Methods The HepG2/ADM was cultured in regular condition with doxorubicin concentrations increasing progressively. The changes of Bcl-2, MDR-1 and TRAILR were quantified in mRNA and protein levels by RT-PCR, Western Blot and immunohistochemistry analysis. Cellular effects of TRAIL in promoting apoptosis in HCC cell line HepG2 and HepG2/ADM were detected by FCM （AnnexinV-FITC-PI assay） after exposure to transfection with pIRES-EGFP-TRAIL. Statistical comparison of means was performed using Student＇s test. Result Expression of death and decoy receptors of HepG2/ADM was not significant different in mRNA and protein levels compared with HepG2, though the HepG2/ADM cells were also resistant to TRAIL alone. Interestingly, the apoptosis rate of the group of TRAIL in HepG2/ADM cells （8.82（0. 37%） is similar with that of the group of TRAIL in HepG2（8.53（0.56%） cells. Conclusion TRAIL has advantage in treating the HepG2/ADM cells.
出处
《肝胆外科杂志》
2005年第4期305-308,共4页
Journal of Hepatobiliary Surgery
关键词
TRAIL
肝癌
TRAIL
Hepatocellular carcinoma
