摘要
目的探讨表皮生长因子受体(EGFR)酪氨酸激酶抑制剂Gefitinib对人肝癌Bel7402细胞株的放射增敏作用及机制。方法实验分四组:实验组(药物Gefitinib+放疗)、药物组、放疗组、对照组(无药物和放疗),采用流式细胞技术分别检测四组作用于肝癌Bel7402细胞株后细胞周期分布状况。细胞免疫化学方法从形态学上观察相关蛋白(Mre11、P53、CDK1)在各实验组的表达情况,Westernblot方法分别检测四组作用肝癌Bel7402细胞株后Mre11、P53、CDK1蛋白的表达情况。结果 (1)流式细胞技术显示:药物组G0/G1期细胞比例略有增长,S期细胞比例减少;放疗组G2/M期细胞比例呈上升趋势,而S期细胞比例下降;实验组G2/M期细胞比例显著增加,G0/G1期及S期细胞均呈现明显下降趋势,且其对S期细胞的比例减少更优于其他三组(P<0.05)。(2)细胞免疫化学法显示Mre11、P53、CDK1蛋白在肝癌Bel7402细胞中均有表达,P53实验组面密度、光密度值与其他三组比较均升高,有显著差异,Mre11、CDK1实验组面密度、光密度值均低于其他三组。(3)Westernblot结果显示,在对肝癌Bel7402细胞株Mre11、CDK1蛋白的表达中,药物组及对照组无明显变化,放疗组及实验组均可使其表达下降,而以实验组对其表达下降的趋势更加显著。实验组与其他组比较P53表达水平明显增高。结论 (1)Gefitinib与射线共同作用于肝癌Bel7402细胞株后放射敏感的G2/M期阻滞增加,放射耐受的S期比例下降。(2)实验组可使Mre11、CDK1蛋白表达下降,实验组下降更明显,实验组与其他组比较:P53表达水平明显增高,Gefitinib可能上调人肝癌Bel7402细胞放射敏感性。
Objective To explore the effection of the inhibitor of EGFR tyrosine kinase Gefitinib for radiosensitization of human hepatocellular carcinoma Bel7402 cell line.Methods Block design:experimental group(drug Gefitinib+radiotherapy);drug group;radiotherapy group;control group.The distribution of cell cycle of the treated human hepatocellular carcinoma Bel7402 cells were detected by flow cytometry.The expression of Mre11 protein,P53 protein and CDK1 protein in different groups by immunocytochemistry were recorded.The expressions of Mre11 protein,P53 protein and CDK1 protein of the treated human hepatocellular carcinoma Bel7402 cells were explored by Western blot.Results (1)Flow cytometry:Drug group:the cell ratio of G0/G1 slightly increased,the cell ratio of S phase decreased;radiotherapy group:the cell ratio of G2/M showed ascendant trend,and the cell ratio of S decreased;experimental group:the cell ratio of G2/M markedly increased,and the cell ratio of G0/G1 and S phase showed obvious downtrend,and the cell ratio of S phase decreased more obviously than drug group and radiotherapy group(P<0.05).(2)Immunocytochemistry:Mre11,P53,CDK1 protein were all expressed in human hepatocellular carcinoma Bel7402 cells.The areal recording density and optical density of P53 protein were significant higher in experimental group than other groups,and the areal recording density and optical density of Mre11 and CDK1 protein were obviously lower in experimental group than the other three groups.(3)The expressions of Mre11 protein of the treated human hepatocellular carcinoma Bel7402 cells in drug group and control group had no obvious change,while decreased in radiotherapy group and experimental group,and decreased most obviously in experimental group(P<0.05).P53 protein and P21 protein were obviously decresed in experimental group than other groups(P<0.05).Conclusions (1)The radiosensitive cell ratio of G2/M of human hepatocellular carcinoma Bel7402 cells increased after treated by Gefitinib combined with radiotherapy,while the radioresistant cell ratio of S phase decreased.(2)The expressions of Mre11 protein of the treated human hepatocellular carcinoma Bel7402 cells in drug group and control group had no obvious change,while decreased in radiotherapy group and experimental group,and decreased most obviously in experimental group,P53 protein and P21 protein were obviously decresed in experimental group than other groups,inferred that Gefitinib increased cell radiation sensitivity.
出处
《中华临床医师杂志（电子版）》
CAS
2012年第19期5843-5847,共5页
Chinese Journal of Clinicians(Electronic Edition)
关键词
肝肿瘤
辐射耐受性
吉非替尼
Liver neoplasms
Radiation tolerance
Gefitinib
