肝细胞生长因子抑制肿瘤细胞凋亡作用研究  

Protection Effect of Hepatocyte Growth Factor on Tumor Cells from Apoptosis and Its Mechanism

作　　者：沈蓉蓉[1] 郑筱娇[1] 赵行[1] 岑东 罗建平 吕建新 裴仁治 滑世轩[4] 

机构地区：[1]温州医学院,浙江省医学遗传学重点实验室,浙江温州325027 [2]浙江省鄞州疾病预防控制中心,浙江宁波315100 [3]浙江省鄞州人民医院血液科,浙江宁波315040 [4]河南省人民医院输血科,郑州450000

出　　处：《中国药学杂志》2012年第18期1478-1482,共5页Chinese Pharmaceutical Journal

基　　金：浙江省医药卫生科技项目(2006A175);宁波市科技计划(2007C10065、2010A610031)

摘　　要：目的探讨肝细胞生长因子(HGF)抑制凋亡诱导剂依托泊苷(etoposide,VP-16)诱导5种肿瘤细胞(B细胞淋巴瘤细胞系Raji、人急性粒细胞白血病细胞系HL-60、宫颈癌细胞系HeLa、前列腺癌细胞系PC-3、非小细胞肺癌细胞系A549)凋亡的作用及其分子机制的研究。方法实验设正常对照组、药物组及肝细胞生长因子保护组,通过CCK-8法观察依托泊苷对5种肿瘤细胞增殖的抑制作用;通过流式细胞术,吖啶橙(AO)荧光染色,瑞氏染色(HE),透射电镜定性及定量观察5种肿瘤细胞的凋亡情况,并进行相关分析。结果 CCK-8法显示依托泊苷可明显抑制Raji、HL-60、PC-3、HeLa、A549等5种细胞增殖的药物浓度分别是100、1、400、200、200μg.mL-1;透射电镜下可观察到典型的凋亡细胞。流式细胞仪检测结果表明,5种细胞的药物组凋亡率均明显高于对照组(P<0.01,P<0.05),其中肝细胞生长因子保护组凋亡率明显低于药物组(P<0.01,P<0.05);吖啶橙染色和HE染色结果表明,正常组细胞形态完好,药物组及肝细胞生长因子保护组凋亡率明显增高(P<0.01,P<0.05),其中肝细胞生长因子保护组凋亡率明显低于给药组(P<0.01,P<0.05)。结论肝细胞生长因子可明显抑制依托泊苷诱导的5种肿瘤细胞凋亡,其作用机制有待进一步研究证实。OBJECTIVE To investigate the protection of hepatocyte growth factor （HGF） on 5 kinds of tumor cells （ B ceil lym- phoma cell line Raji, human acute myeloid leukemia cell line HL-60, cervical cancer cell line HeLa, prostate cancer cell line PC-3, and non-small cell lung cancer cell line A549） from apoptosis induced by etoposide （VP-16）. METHODS Normal control group, drug group, and HGF protection group were set. CCK-8 assay was used to measure proliferation inhibition on 5 kinds of tumor cells by VP-16. Quantitative and qualitative analysis on 5 kinds of tumor cells was performed through acridine orange （AO） fluorescent stai- ning, flow cytometry, HE staining, and transmission electron microscopy. RESULTS CCK-8 assay revealed that the concentrations of VP-16, which can significantly inhibit the proliferation of Raji, HL-60, PC-3, HeLa, and A549 cell lines, were 100, 1 , 400, 200, and 200 I^g ＂ mL -~ , respectively. The typical morphologic changes of cell apoptosis were observed under transmission electronic micro- scope. Flow cytometry showed that the apoptotic rates of tumor cells in the drug groups were significantly higher than those in noruaal control group （ P 〈 0. 01, P 〈 0.05 ） and in HGF protection groups （ P 〈 0. 01, P 〈 0.05 ）. AO and HE staining revealed that ceils in normal control group appeared to have regular cell morphology, but the cells apoptotic rates in the drug groups were significantly higher than those in normal control groups （P 〈0. 01, P 〈0. 05） and in HGF protection groups （P 〈0. 01, P 〈0. 05）. CONCLUSION HGF can significantly protect 5 kinds of tumor ceils from apoptosis induced by VP-16. The mechanism need further investigation.

关 键 词：肝细胞生长因子 依托泊苷 凋亡 流式细胞术 吖啶橙染色 肿瘤 

分 类 号：R965[医药卫生—药理学]