沉默Glypican-3基因增强人肝癌细胞HepG2对5-FU致凋亡作用的敏感性  

Silencing Gene of Glypican-3 Increases the Sensitivity to 5-FU-induced Apoptosis in Hepatocellular Carcinoma Cells HepG2

作　　者：王孝茹[1] 董红霖[1] 苏继荣[1] 胡杰[1] 马瑞[1] 薛利利[1] 

机构地区：[1]山西医科大学第二医院普外科,山西太原030001

出　　处：《中国现代医生》2011年第11期6-8,18,共4页China Modern Doctor

摘　　要：目的沉默人肝癌细胞HepG2的GPC3基因表达,观察其对5-氟尿嘧啶(5-FU)的药物敏感性及细胞凋亡情况的影响。方法①构建干扰GPC3的shRNA载体,将其转染至肝癌细胞,并设立对照组。②Western blot测定GPC3蛋白表达情况。③利用MTT法测定转染后肝癌细胞对5-FU的药物敏感性。④采用流式细胞技术观察转染后肝癌细胞在5-FU作用下的细胞凋亡情况。结果转染shRNA的肝癌细胞其GPC3表达阴性;其对5-FU的药物敏感性增加(P<0.05);细胞凋亡较对照组增加(P<0.05)。结论 GPC3的高表达在肝癌细胞中起到抗凋亡作用,而下调GPC3可作为一个潜在的靶目标,增加肝癌细胞对5-FU诱导细胞凋亡的敏感性。Objective Silencing gene of glypican-3 was performed in HCC cells HepG2, investigate the sensitivity to 5-FU and the effect of apoptosis induced by 5-FU. Methods We generated a hepatocellular carcinoma cell line stably transfected with plasmids encompassing shRNA targeting GPC3,and control was used. Western blot analysis was performed to assess the expression of GPC3. The sensitivity to 5-FU in HepG2 was evaluated by M3T assay. Flow cytometer was used to observe the effect of apoptosis induced by 5-FU. Results The expression of GPC3 was negative in HepG2 cells transfected with shRNA. HCC cell line with silenced GPC3 expression was more sensitive to 5-FU （P〈O.O5）;and 5-FU-treated shGPC3 cells showed a signi？cant increase in apoptosis（P〈O.05） Conclusion Increased GPC3 expression in HCC cell line may play a protective role to against apoptosis, and the down regulation of GPC3 may be a potential target to increase sensitivity to 5-FU-induced apoptosis in hepatocellular carcinoma cells.

关 键 词：肝细胞癌 GLYPICAN-3 SHRNA 5-氟尿嘧啶 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]