联合中药消积煎剂综合治疗对顺铂耐药卵巢癌体外作用的实验研究

Effect of a Combined Treated with Xiaoji Decoction about Hyperthermia and Cisplatin in Dealing with Cisplatin-resistant Human Ovarian Cancer Cell

目的:进行体外药物敏感试验,应用综合治疗方案提高化疗药物的敏感性,探索克服卵巢恶性肿瘤抗药性,提高患者生存率的有效途径。方法:将消积煎剂(0.5、5、50mg/mL)、热疗(42℃)和顺铂(6、12、24μg/mL)分别单独以及联合作用于体外培养的人卵巢癌CAOV3/cDDP细胞株。采用MTT(methyl thiazolyl tetrazolium)法检测各实验组人卵巢癌耐药细胞株CAOV3/cDDP的生长抑制率;采用集落形成实验观察各实验组对人卵巢癌耐药细胞株CAOV3/cDDP的诱导分化作用;应用光镜观察实验处理前后人卵巢癌CAOV3/cDDP细胞形态和结构的变化,确定凋亡细胞,评价各组细胞凋亡程度。结果:消积煎剂、热疗和顺铂3种疗法同时应用,结果显示为很明显的协同作用(P<0.01)。形态学观察可看到,实验处理后人卵巢癌CAOV3/cDDP细胞出现典型的凋亡现象。结论:消积煎剂、热疗和顺铂联合应用可逆转肿瘤细胞对顺铂的耐药性,有很好的临床应用前景。

Objective ： The objective of this study to evaluate the effect of a concomitant treatment with Xiaofi Decoction, hyperthermia and cisplatin in drug - resistant human ovarian cancer cell line CAOV3/cDDP. Methods ： CAOV3/ cDDP cell line were treated by Xiaofi Decoction （0.5mg/mL, 5mg/mL,50mg/mL）, hyperthermia （42℃） or cisplatin （6μg/mL,12μg/mL,24μg/mL） alone, or by the combination of them. The growth inhibiting rates to human ovarian cancer cell line CAOV3/cDDP were determined by methyl thiazolyl tetrazolium （MTT） method. Signs of induction differentiation were observed by colony forming units assay, respectively. Light microscopy was applied to examine the changes in morphology and structure of CAOV3/cDDP cell line, in order to evaluate cell apoptosis. Results ：It was found that compared with various concentrations of cisplatin, hyperthermia at 42℃ for 60 min with cisplatin markedly inhibited the cell growth （ P 〈 0.01 ）. When all treatments were given concurrently, the response was greater than calculated response of all three individual treatments, indicating a synergistic interaction （ P 〈 0.01 ）. Morphological observation indicated that Xiaofi Decoction inducing CAOV3/cDDP death characterized by apoptotic phenotype. Conclusion ： The combination of hyperthermia, cisplatin and Xiaoji Decoction is not only a good combination for potential clinical application, but also an effect way of dealing with cisplatin resistance.