紫杉醇诱导的卵巢癌耐药细胞株的生物学特性及其保存

发布时间：2018-04-28 06:39

本文选题：紫杉醇 + 耐药　；参考：《肿瘤防治研究》2015年04期

【摘要】：目的研究紫杉醇诱导的卵巢癌耐药细胞株的生物学特性,探索耐药细胞的长期保存条件,以建立稳定的体外实验模型。方法选用卵巢癌化疗敏感细胞株SKOV3,分别采用大剂量冲击和小剂量浓度递增诱导建立耐药的卵巢癌细胞株SKOV3/TAX300、SKOV3/TAX30。四甲基偶氮唑盐(MTT)法检测耐药细胞对化疗药物紫杉醇的IC50;比较耐药细胞和敏感细胞的细胞平板克隆形成、生长曲线倍增时间的差异;荧光定量PCR检测敏感细胞和耐药细胞中耐药相关基因的表达;比较无药冻存和加药冻存的方式对耐药细胞的保存效果。结果耐药细胞SKOV3/TAX300,耐药指数为4.60,耐药细胞SKOV3/TAX30,耐药指数为51.31;与敏感细胞相比,耐药细胞体积增大,形态不规则,倍增时间延长。多药耐药基因1(MDR1)在两种耐药细胞中高表达,肺耐药相关蛋白(LRP)、蛋白激酶Cα(PRKCA)基因仅在SKOV3/TAX300中的表达增高,BCL-2样1基因(BCL2L1)在SKOV3/TAX30中低表达。耐药细胞冻存在含有药物的条件下,有利于耐药性的保持。结论大剂量冲击和小剂量浓度递增两种方法诱导的耐药细胞为研究紫杉醇耐药机制建立了良好的体外模型,两种耐药细胞SKOV3/TAX300和SKOV3/TAX30具有不同的生物学特性。
[Abstract]:Objective to study the biological characteristics of paclitaxel induced ovarian cancer cell lines and to explore the long-term preservation conditions of drug resistant cells in order to establish a stable experimental model in vitro. Methods the chemotherapeutic sensitive cell line SKOV3 of ovarian cancer was selected and the drug resistant ovarian cancer cell line SKOV3/TAX30 was induced by high dose impact and low dose concentration increase respectively. 0, SKOV3/TAX30. four methyl azazolium salt (MTT) assay was used to detect the IC50 of chemotherapeutic drug paclitaxel, the formation of cell plate clone of drug resistant cells and sensitive cells, the difference of multiplication time of growth curve, the expression of drug resistance related genes in sensitive and drug-resistant cells by fluorescence quantitative PCR, and the comparison of drug free cryopreservation and drug freeze. The resistance cell SKOV3/TAX300, the resistance index was 4.60, the resistance cell SKOV3/TAX30, the resistance index was 51.31, the volume of the drug resistant cells, the irregular shape, the doubling time, and the high expression of multidrug resistance gene 1 (MDR1) in two resistant cells and lung resistance related eggs. White (LRP), protein kinase C alpha (PRKCA) gene only increased in SKOV3/TAX300, BCL-2 like 1 gene (BCL2L1) was low in SKOV3/TAX30. Drug-resistant cells were frozen in the presence of drugs and were beneficial to the retention of drug resistance. Conclusion two methods induced by large dose impact and low dose concentration were used to study taxol tolerance The drug mechanism established a good in vitro model, and the two kinds of drug-resistant cells SKOV3/TAX300 and SKOV3/TAX30 have different biological characteristics.

【作者单位】：首都医科大学附属北京妇产医院妇瘤科;首都医科大学附属北京世纪坛医院妇产科;
【基金】：首都医科大学基础临床合作研究基金(12JL64)
【分类号】：R737.31

【参考文献】