放射联合Sorafenib对肝癌细胞的不同时效作用  

Schedule-dependant in Vitro Anti-tumor Effect of Radiation Combined with Sorafenib in Hepatocellular Carcinoma

作　　者：李巧巧[1] 刘孟忠[1] 王莉[2] 李锦清[3] 

机构地区：[1]华南肿瘤学国家重点实验室∥中山大学肿瘤防治中心放疗科,广东广州510060 [2]华南肿瘤学国家重点实验室∥中山大学肿瘤防治中心研究所,广东广州510060 [3]华南肿瘤学国家重点实验室∥中山大学肿瘤防治中心肝胆科,广东广州510060

出　　处：《中山大学学报（医学科学版）》2010年第5期646-651,680,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家"十一五"攻关项目(2006BA102A04)

摘　　要：【目的】在人肝癌细胞中观察放射联合Sorafenib对细胞生长的作用,并探讨其作用途径。【方法】选择人肝癌SMMC-7721和BEL-7402细胞株,分为单纯放射组(IR)、放射前30min联合Sorafenib组(IR+S-pre)和放射24h后联合Sorafenib组(IR+S-post)。细胞克隆形成实验检测照射后细胞克隆形成率,绘制细胞存活曲线并计算放射增敏比;细胞增殖抑制实验检测6Gy照射后第1、2、3、4、5、6d细胞增殖情况;免疫荧光染色观察照射后DNA损伤阳性细胞比例;流式细胞仪检测放射后细胞凋亡比例。【结果】细胞克隆形成实验结果显示,IR+S-pre放射后细胞克隆形成增加,放射增敏比在SMMC-7721和BEL-7402细胞中分别为0.78和0.88;而IR+S-post放射后细胞克隆形成减少,放射增敏比在SMMC-7721和BEL-7402细胞中分别为1.43和1.23。细胞增殖抑制实验显示IR+S-pre与IR放射后细胞增殖抑制率相似,而IR+S-post放射后细胞增殖抑制率明显提高。Sorafenib在SMMC-7721与BEL-7402中均有诱导细胞凋亡作用。IR+S-pre在放射后24h细胞凋亡比例明显增高[IRvsIR+S-pre:(6.1±1.0)%vs(18.3±2.0)%(SMMC-7721),(8.2±2.1)%vs(17.0±2.4)%(BEL-7402),P<0.001]。IR+S-post在放射后48h细胞凋亡比例明显增高,[IRvsIR+S-post分为(6.9±2.0)%vs(15.9±1.8)%(SMCC-7721),(8.0±1.5)%vs(14.2±2.5)%(BEL-7402),P<0.05]。放射后30min,IR+S-pre与IR的DNA受损阳性细胞比例无明显差异。放射后6hIR+S-pre的DNA受损阳性细胞残留比例明显下降[IRvsIR+S-pre:(59.9±2.4)%vs(23.8±2.9)%(SMMC-7721),(46.4±3.8)%vs(25.0±3.0)%(BEL-7402),P<0.001]。【结论】放射联合Sorafenib对肝癌细胞作用具有明显时效性,放射24h后联合Sorafenib增加放射引起的细胞克隆性生长抑制和增殖抑制,而放射前30min联合Sorafenib作用相反。Objective The effect of radiation combined with Sorafenib in hepatocellular carcinoma cells（HCC） and the mechanism of combination was investigated.Methods HCC lines SMMC-7721 and BEL-7402 were studied in 3 groups respectively：irradiation only（IR group）,radiation with Sorafneib 30 min pre-irradiaiton（IR ＋ S-pre group） and radiation with Sorafenib 24 h post-irradiaiton（IR ＋ S-post group）.The cells were irradiated in different dose with or without Sorafenib.The different groups were compared according to the survival curves processed by clonogenic assay and the MTS assay in different days（1,2,3,4,5,and 6） after irradiated by 6 Gy.Then the tumor cells were irradiated by a dose of 6 Gy and their DNA damage after irradiation were determined respectively through γ-H2AX focus by immunoflurescence.The apoptosis cells were determined by flow cytometry.Result The HCC cells had more colonies and similar proliferation inhibitor by IR ＋ S-pre,and less colonies and higher proliferation inhibitor by IR ＋ S-post compared with IR.The sensitivity enhance ratios（SERSF2） were 0.78 and 0.88 in IR＋ S-pre group in SMMC-7721 and BEL-7402 respectively,and 1.43 and 1.23 in IR ＋ S-post group in SMMC-7721 and BEL-7402 respectively.Sorafenib increased apoptosis in SMMC-7721 and BEL-7402.The IR ＋ S-pre group had more apoptosis cells 24 h after radiation [IR vs IR＋S-pre：（6.1 ± 1.0）% vs（18.3 ± 2.0）% in SMMC-7721 and（8.2 ± 2.1）% vs（17.0 ± 2.4）% in BEL7402,P0.001].The IR ＋ S-post group had more apoptotic cells 48 hours after radiation [IR vs IR ＋ S-post：（6.9 ± 2.0）% vs（15.9 ± 1.8）% in SMCC-7721 and（8.0 ± 1.5）% vs（14.2 ± 2.5）% in BEL-7402,P0.05].The proportions of DNA damage positive cells were similar 30 min after irradiation,but less in IR ＋ S-pre group 6 h after irradiation（IR vs IR ＋ S-pre：（59.9 ± 2.4）% vs（23.8 ± 2.9）% in SMMC-7721 and（46.4 ± 3.8）% vs（25.0 ± 3.0）% in BEL-7402,P0.001） compared with IR group.Conclusion Radiatio

关 键 词：肝癌 放射 索拉非尼 凋亡 DNA损伤修复 

分 类 号：R73-36[医药卫生—肿瘤]