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羟基喜树碱和topotecan对人卵巢癌细胞株体外作用的研究 |
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th of SKOV3 and CAOV3 was dose-dependent. 50% inhibiting concentration (IC50) of HCPT and topotecan to SKOV3 was 72 ng/ml and 160 ng/ml, respectively; IC10 of HCPT and topotecan to CAOV3 was 141 ng/ml and 12 ng/ml, respectively. The inhibition of HCPT and topotecan to clone forming rate of two cell lines was significant. The growing curves of SKOV3 and CAOV3 treated by HCPT and topotecan were significantly lower than controls. HCPT and topotecan of 5 ng/ml can elongate the double time of SKOV3 for 49.00 and 1.75 times, respectively. They can elongate the double time of CAOV3 for 2.77 and 1.94 times, respectively. After treated by HCPT or topotecan, SKOV3 generated DNA Ladder. Positive rates of TUNEL of SKOV3 and CAOV3 were elevated by HCPT and topotecan notably. HCPT and topotecan did not affect the expression of c-myc, bcl-2, fas of SKOV3 and CAOV3. Observed by electronic microscope, heterochromatin of SKOV3 and CAOV3 coagulated after treated by HCPT. Micro villi of SKOV3 and CAOV3 were damaged, and some vacuoles appeared in the cytoplasm. Conclusions HCPT and topotecan can inhibit growth of SKOV3 and CAOV3 significantly. HCPT and topotecan can induce apoptosis of SKOV3 and CAOV3.
【Key words】 Lamptothecin Topotecan Ovarian neoplasms
10-羟基喜树碱（羟基喜树碱）主要在我国应用于肝癌、胃肠道肿瘤、膀胱癌的治疗；而9-二甲基氨基-10-羟基喜树碱（topotecan）主要在国外应用于肺癌、卵巢癌，尤其是对顺铂治疗失败或复发的卵巢癌患者，有较好的治疗作用［1,2］。本研究以卵巢癌细胞株SKOV3和CAOV3为对象，研究羟基喜树碱和topotecan对这两种卵巢癌细胞株体外生长的影响及诱导这两种细胞凋亡的情况，并将这两种药物与顺铂进行了对比研究。现报道如下。
材料和方法
一、材料
1．卵巢癌细胞株：（1）SKOV3，来源于卵巢癌患者腹水，美国G.Trempe于1973年建立；（2）CAOV3，来源于卵巢癌患者肿瘤组织，为美国J.Frog于1976年建立。两种细胞株均由美国国立癌症研究所赠送我院妇科肿瘤实验室，本实验中发现细胞株CAOV3为顺铂耐药株。
2．试剂：四甲基偶氮唑蓝（MTT）为美国Sigma公司生产；原位细胞凋亡检测试剂盒购自德国Boehringer-Mannheim公司；抗c-myc蛋白抗体、抗bcl-2蛋白抗体、抗fas蛋白抗体，均为美国Santa Cruz公司生产，购自北京中山公司。
二、方法
（一）测定羟基喜树碱和topotecan对SKOV3和CAOV3生长影响的实验
1. MTT显微培养实验：以无菌操作技术分别配制浓度为0.244～64 000 ng/ml的羟基喜树碱、topotecan和顺铂的培养基，药物浓度以2倍递增。用以上浓度的含药培养基孵育细胞，分别在24小时和48小时时进行MTT快速比色，在酶标仪上一页 [1] [2] [3] 下一页
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