化疗药物杀伤原代胃癌细胞效应与P-糖蛋白和GST-π表达的关系  

Antitumor Effects of Chemotherapeutic Drugs on Fresh Human Gastric Cancer Cells and Their Relationships to Expressions of P-glycoprotein and Glutathione S Transferase-π

作　　者：张俊[1] 计骏[1] 纪玉宝[1] 袁菲[2] 刘炳亚[1] 朱正纲[1] 林言箴[1] 

机构地区：[1]上海第二医科大学附属瑞金医院外科上海消化外科研究所,上海200025 [2]上海第二医科大学附属瑞金医院病理科,上海200025

出　　处：《癌症》2005年第4期432-437,共6页Chinese Journal of Cancer

基　　金：上海市医学发展基金(No.983008)~~

摘　　要：背景与目的:化疗耐药是影响进展期胃癌疗效的主要原因之一。本研究旨在评价不同化疗药物对原代胃癌细胞的体外杀伤效应及其诱导凋亡能力,同时研究上述效应与胃癌组织P鄄糖蛋白(P鄄gp)和谷胱甘肽S转移酶π(GST鄄π)表达的关系。方法:39例分离纯化后的人新鲜胃癌细胞,分别暴露于血浆峰浓度的5鄄氟尿嘧啶(5鄄FU)、顺铂(DDP)、丝裂霉素(MMC)、阿霉素(ADM)及羟基喜树碱(HCPT)。用台盼蓝染色法和MTT法检测经上述药物作用后肿瘤细胞活力及代谢活性的变化;用原位末端转移酶标记法(TdT法)检测胃癌细胞凋亡率;并用免疫组化染色检测胃癌组织中GST鄄π和P鄄gp的表达。结果:经化疗药物作用后,体外培养的胃癌细胞出现不同程度的形态学改变、代谢活性下降及细胞凋亡。不同个体肿瘤细胞对不同化疗药物的敏感性不同。MTT结果提示,所测药物对胃癌细胞的平均抑制率由高到低依次为MMC[(38.6±7.7)%],DDP[(38.1±8.8)%],5鄄FU[(37.8±10.3)%],ADM[(31.9±10.4)%],HCPT[(29.9±10.2)%]。MMC、DDP及5鄄FU的平均抑制率明显高于HCPT及ADM(P<0.01)。TdT结果提示,诱导胃癌细胞凋亡率由高到低的药物依次为DDP[(32.1±7.7)%],5鄄FU[(31.1±8.8)%],MMC[(29.8±6.3)%],HCPT[(21.9±7.4)%],ADM[(19.9±7.4)%]。本组病例GST鄄π和P鄄gp的?BACKGROUND & OBJECTIVE: Drug-resistance is a major factor of influenc ing treatment efficacy of advanced gastric cancer. This study was to evaluate in vitro antitumor effects of different chemotherapeutic drugs on fresh human gast ric cancer cells, and explore their relationships with expressions of P-glycopro tein (P-gp), and glutathione S transferase -π (GST-π) in human gastric cancer tissue. METHODS: A total of 39 specimens of highly purified gastric cancer cells were separately exposed to 5-fluorouracil (5-FU), cisplatin (DDP), mitomycin C (MMC), adriamycin (ADM), and hydroxycamptothecin (HCPT). Inhibitory rate of cell s was detected by MTT assay. Metabolic activity of cells was detected by trypan blue exclusive assay. Cell apoptosis was detected by in situ terminal deoxynucle otidyl transferase assay (TdT assay). Expressions of GST-π and P-gp in gastric cancer tissues were detected by immunohistochemistry. RESULTS: After exposure to antitumor drugs, morphologic changes, decrease of metabolic activity, and apopt osis were appeared in gastric cancer cells. Inhibitory rates of cancer cells exp osed to MMC, DDP, 5-FU were significantly higher than those of cells exposed to ADM, and HCPT <(38.6±7.7)%, (38.1±8.8)%, and (37.8±10.3)% vs. (31.9±10.4)%, and (29.7±10.2)%, P < 0.01>. Apoptosis rate of cancer cells exposed to CDDP, 5-FU, and MMC were significantly higher than those of cells exposed to HCPT, and ADM <(32.1±7.7)%, (31.1±8.8)%, and (29.8±6.3)% vs. (21.9±7.4)%, and (19.9± 7.4)%, P < 0.05>. Positive rate of GST-π was 66.7%(26/39), that of P-gp was 59 .0%(23/39). GST-π positive cells showed resistance to DDP and MMC, P-gp positiv e cells showed resistance to ADM and HCPT. CONCLUSIONS: Overexpressions of P-gp and GST-π might contribute to drug-resistance of tumor. Detection of GST-π and P-gp, together with MTT chemosensitivity test, might be useful for selecting mo re effective chemotherapeutic drugs.

关 键 词：胃肿瘤 抗癌药物 谷胱甘肽S转移酶Π P糖蛋白 

分 类 号：R735.2[医药卫生—肿瘤]