细胞凋亡检测用于肿瘤细胞株对化疗敏感性的研究

目的 评价Annexin-V荧光标记FACScan法、TUNEL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）、1／10PPC、1／5PPC、5PPC、10PPC与结肠腺癌LoVo和Ls-174-t细胞温育24及48h，用FACScan法、TUNEL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 LoVo与Ls-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，TUNEL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＜0．05）。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、选用的方法。

STUDY OF CHEMOTHERAPYSENSITIVITY IN TUMOR CELL LINES ASSESSED BY APOPTOSIS

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.