顺铂和阿霉素诱导肝细胞肝癌细胞凋亡阈值研究

Study of apoptotic thershold of cis-diamminedichloroplatinum and adriamycin on hepatocellular carcinoma

目的 :了解抗癌药物顺铂 (CDDP)、阿霉素 (ADM)诱导肝细胞肝癌 (HCC)的细胞凋亡阈值。方法 :采用肿瘤细胞原代培养技术、活细胞萤光染色法和流式细胞仪定量分析。结果 :顺铂或阿霉素与细胞共同培养后 ,荧光显微镜下见正常细胞核呈弥散均匀荧光 ,凋亡细胞核内颗粒状荧光。随抗癌药物 (ADM、CDDP)剂量的增加 ,诱导人肝癌细胞的细胞凋亡率也增加 ,但以ADM 1.0 μg/mL和 2 0 μg/mL及CDDP 1 5 μg/mL和 3 0 μg/mL作用明显。ADM 2 0 μg/mL可导致HCC细胞凋亡 75 % ,CDDP 3 0 μg/mL可致HCC细胞凋亡 75 %。 2 5例未经治疗的HCC细胞和HepG - 2细胞的ADM、CDDP敏感性检测表明 ,ADM的凋亡阈值为 1 0 μg/mL ,CDDP的凋亡阈值为 1 5 μg/mL。临床凋亡敏感剂量分别为ADM 2 0mg/m2 ,CDDP 30mg/m2 。结论 :本文获得的ADM和CD

AIM:To study the thershold of hepatocellular carcinoma (HCC) apoptosis induced by adriamycin (ADM) and cis-diamminedichloroplatinum(CDDP). METHODS: Using primary human hepatocellular carcinoma culture, immunofluorescence staining of Hoechst 33 258 in HCC cells ,and flow cytometric assay. RESULTS:24 h after HCC cells were cultured with ADM or CDDP, it were found there were dispersive fluorescences in normal cells nuclei, and compact particulate fluorescences in apoptosis cells nuclei by immunofluorescence staiming of Hoechst 33 258. The rate of HCC cells apoptosis was dependent on doses of ADM and CDDP. HCC cells apoptotic threshold were determined, i.e.ADM was 1.0 μg/mL and CDDP was 1.5μg/mL (The clinical apoptotic sensitive dosages were ADM 20 mg/m2 and CDDP 30 mg/m2. CONCLUSION: HCC cells apototic threshold of ADM and CDDP were efficient in clinical chemotherapy.