细胞凋亡检测用于肿瘤细胞株对化疗敏感性的研究

STUDY OF CHEMOTHERAPY SENSITIVITY IN TUMOR CELL LINES ASSESSED BY APOPTOSIS

目的 评价 Annexin- V荧光标记 FACScan法、TU NEL法细胞凋亡检测在肿瘤细胞株化疗药物敏感性研究中的应用。方法 将 DDP、MMC、5 - FU、EPI按血浆峰浓度 (PPC)、1/10 PPC、1/5 PPC、5 PPC、10 PPC与结肠腺癌L o Vo和 L s- 174- t细胞温育 2 4及 48h,用 FACScan 法、TU NEL法检测细胞凋亡 ,用 MTT比色法检测细胞生长抑制率 ,并提取 DNA进行琼脂糖凝胶电泳。结果 L o Vo与L s- 174- t细胞经化疗药物诱导 48h后 ,FACScan检测的细胞凋亡率在 PPC时最高 ,TU NEL 法检测的凋亡率与 MTT法检的细胞增殖抑制率呈正的直线相关 (P<0 .0 5 )。结论 Annexin- V荧光标记 FACScan法检测细胞凋亡既能优选出敏感的药物 ,又能寻找合适的药物浓度 ,是目前优选有效化疗药物种类及剂量的一个可探索。

Objective To study chemotherapy sensitivity and apoptosis in tumor cell line.Methods Two different human colon carcinoma cell lines, LoVo and Ls 174 t were incubated with DDP,MMC,5 FU,EPI at various peak plasma concentrations (PPC), 1/10PPC,1/5PPC,5PPC and 10 PPC. Apoptosis was detected by FACScan and TUNEL technique after 24 hours and 48 hous. Cell growth inhibition rates were assessed by MTT, DNA ladder were examined by agarose electrophoresis.Results At 48 hours, the highest cellular apoptosis rates were observed with PPC which was assessed by FACScan technique. There was a positive linear correlation between apoptosis assessed by TUNEL technique and growth inhibition rates of cells determined by MMT( P <0 05).Conclusion Using Annexin V Fluos staining FACScan technique to assess apoptosis of tumor cells is an exploring and useful method which can be used to choose both kinds and doses of chemotherapeutic drugs.