黄连素逆转结肠癌细胞株HCT-8/VCR多药耐药及其与P-gp功能变化相关性的探索性研究

目的观察黄连素对结肠癌细胞株HCT-8/VCR耐药性的逆转作用,并从P-gp功能变化的角度分析黄连素逆转细胞耐药性的机制。方法以人结肠癌细胞株HCT-8/VCR为观察对象,选用无干预措施、不同浓度黄连素处理(实验组)、维拉帕米处理组(阳性对照组)3种处理方法,采用MTT法测定不同浓度的长春新碱对3组细胞的抑制率,并计算耐药逆转倍数。采用微孔板荧光分光广度计测定3种处理方法细胞种罗丹明123的荧光强度,分析黄连素对于P-gp转运功能的影响。结果 MTT实验结果显示,不同浓度的长春新碱对于肿瘤细胞的具有抑制作用,且这种抑制作用与药物浓度呈正相关,药物对于细胞IC50为(18.31±0.57)μg/mL。加入黄连素处理细胞后,细胞对于药物的敏感性增加,黄连素的逆转倍数为3.57。罗丹明123排出实验结果显示,细胞对于罗丹明123的排出能力随着黄连素浓度的升高而增加。细胞酶联免疫吸附实验结果显示不同浓度的黄连素处理24h后,细胞的P-gp表达量均呈现明显降低。结论黄连素能够提升结肠癌细胞对化疗药物的敏感性,且这种机制可以通过抑制P-gp的表达而实验降低药物的外排而实现。

The Exploratory Research on the Berberine Reverse the HCT-8/VCR Multidrug Resistance and Its Relationship with P-gp Functional Variance

Objective To explore the effect of berberine on reversing the HCT-8/VCR multidrug resistance and the relationship with P-gp functional variance.Method The HCT-8/VCR were treated with different concentrations of berberine（experimental group）,verapamil（positive group）,non-intervention（control group）.The MTT method served to determine the inhibition rate of different concentration s of vincristine on the three groups/ The multiple drug resistance reversal was caculated.The light intensity of rhodamine 123 was measured to evaluate the effect of berberine on the P-gp function.Results MTT results showed that different concentration of vincristine can inhibited the growth of tumor cells with IC50（18.31±0.57） μg/mL.The inhibition was positively correlated with drug concentration.After the treatment of berberine,the sensitivity of cells to chemotherapy was increased,with the reverse rate 3.57.The capacity for the Rhodamine 123 can be increased with the concentration of berberine increased.The results of ELIZA showed that the expression of P-gp can be reduced with the treatment of berberine for 24 hours.Conclusions The treatment of berberine can increase the HCT-8/VCR sensitivity to the chemotherapy drugs,which is mediated by decreasing the expression of P-gp.