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目的以卡铂(CBP)和平阳霉素(PYM)为诱导物,构建耐药细胞株Tca8113/CBP和Tca8113/PYM,再通过银杏酸(GAs)与化疗药物联用探讨银杏酸对耐药细胞多药耐药(MDR)的影响。方法免疫组织化学检测P-糖蛋白(P-gp)的表达,MTT法确定耐药细胞的耐药指数;不同质量浓度的GAs作用于耐药细胞及亲本细胞,通过MTT法检测对它们的增殖抑制效应,确定GAs的无细胞毒性质量浓度并观察GAs对耐药细胞的逆转作用;GAs与耐药细胞联用诱导细胞一段时间后,再次通过MTT法检测其耐药指数。结果免疫组织化学结果显示,耐药细胞的P-gp阳性表达率明显高于亲本细胞,MTT法确定GAs的无细胞毒性质量浓度为10μg.mL-1;GAs对Tca8113/CBP和Tca8113/PYM细胞的逆转倍数分别是2.94和2.43,与GAs联用前Tca8113/CBP和Tca8113/PYM细胞的耐药指数分别是3.24和11.9,联用后的耐药指数分别是2.18和4.43。结论本实验成功诱导出了耐药细胞株Tca8113/CBP和Tca8113/PYM,并将银杏酸与化疗药物联用,进一步证实了两者的共用能够增强对Tca8113/CBP和Tca8113/PYM细胞的增殖抑制作用,无细胞毒质量浓度的GAs对Tca8113/CBP和Tca8113/PYM细胞的耐药性有部分逆转作用,且共用一段时间后耐药细胞的MDR水平有所下降。
OBJECTIVE: To construct drug-resistant cell lines Tca8113 / CBP and Tca8113 / PYM by inducing carboplatin (CBP) and bleomycin (PYM), and then to explore the effect of ginkgolic acid on chemoresistance cells Multidrug resistance (MDR) effects. Methods The expression of P-glycoprotein (P-gp) was detected by immunohistochemistry and the drug resistance index of drug-resistant cells was determined by MTT assay. GAs with different concentrations were applied to drug-resistant cells and parental cells, Inhibitory effect of GAs was determined to determine the cytotoxicity of GAs and to observe the reversal effect of GAs on drug-resistant cells. After treated with GAs and drug-resistant cells for a period of time, the resistance index of GAs was tested again by MTT assay. Results Immunohistochemical results showed that the positive expression rate of P-gp in drug-resistant cells was significantly higher than that in parental cells. The cytotoxicity of GAs was determined as 10μg.mL-1 by MTT assay. The effect of GAs on Tca8113 / CBP and Tca8113 / PYM cells Were 2.94 and 2.43, respectively. The resistance index of Tca8113 / CBP and Tca8113 / PYM before combination with GAs were 3.24 and 11.9, respectively, and the combined resistance index was 2.18 and 4.43 respectively. CONCLUSIONS: Tca8113 / CBP and Tca8113 / PYM were successfully induced in this experiment, and the combination of ginkgolic acid and chemotherapeutic agents further confirmed that the combination of the two could enhance the proliferation inhibition of Tca8113 / CBP and Tca8113 / PYM cells The cytotoxicity of GAs on Tca8113 / CBP and Tca8113 / PYM cells partially reversed, and the MDR levels of drug-resistant cells decreased after a certain period of time.