人参皂苷Rg3逆转人胆管癌细胞多药耐药性的实验研究

Experimental Research on Reversal of Multidrug Resistance in Human Cholangiocarcinoma Cell by Ginsenoside Rg3

目的探讨人参皂苷Rg3逆转人胆管癌多药耐药细胞株QBC939/ADM的耐药性。方法人胆管癌多药耐药细胞株QBC939/ADM的建立,主要采用阿霉素(ADM)持续接触浓度递增法,必要时结合高低浓度交替法诱导人胆管癌细胞QBC939产生多药耐药性(MDR)。将三种不同浓度的化疗药物,即环磷酰胺(CTX)、丝裂霉素(MMC)、5-氟尿嘧啶(5-FU)分别作用于QBC939及QBC939/ADM,用CCK-8法检测不同化疗药物的细胞毒作用,流式细胞术检测细胞凋亡率,RT-PCR法检测MDR基因的表达水平,将得到的数据运用统计学分析,并得出结果,从而证明人胆管癌多药耐药细胞株QBC939/ADM的建立。以中药人参的有效成分人参皂苷Rg3作为MDR逆转剂,逆转QBC939/ADM的MDR,用CCK-8法、流式细胞术、RT-PCR法检测其逆转作用。结果CCK-8法检测化疗药物作用于对照组和实验组的细胞毒作用,结果显示:实验组细胞的OD值(吸光度)明显低于对照组,P <0. 05,差异有统计学意义;流式细胞仪检测化疗药物作用于对照组和实验组的细胞凋亡率:实验组的细胞凋亡率明显高于对照组,P <0. 05,差异有统计学意义; RT-PCR法测定MDR基因的表达,结果显示实验组MDR基因的表达明显低于QBC939/ADM,P <0. 05,差异有统计学意义。结论人胆管癌多药耐药细胞株QBC939/ADM成功建立,并初步证明中药人参的有效成分人参皂苷Rg3能有效逆转其MDR,但其逆转机制尚未研究,需进一步探索。

Objective To investigate the effect of ginsenoside Rg3 on multidrug resistance of human cholangiocarcinoma cell line QBC939/ADM. Methods Establishment of multidrug resistant human cholangiocarcinoma cell line QBC939/ADM,mainly by adriamycin( ADM)concentration increasing continuous contact method,with high and low concentration alternating method induced by bile duct cancer cells QBC939 multidrug resistance( MDR) when necessary. The three different concentrations of chemotherapeutic drugs,namely Cyclophosphamide( CTX),Mitomycin( MMC),5-Fluorouracil( 5-FU) respectively for QBC939 and QBC939/ADM,with CCK-8 method to detect the cytotoxic effect of chemotherapeutic agents,cell apoptosis was detected by flow cytometry,M RT-PCR detection method The expression level of DR gene,the resulting data using statistical analysis, and obtains the result, which proves that establishment of multidrug resistant human cholangiocarcinoma cell line QBC939/ADM. The effective constituents of ginsenoside Rg3 in ginseng as MDR reversal agents,reverse QBC939/ADM MDR,by CCK-8 method,flow cytometry,detection of the reversal effect the method of RT-PCR. Results CCK-8 method was used to detect the cytotoxicity of chemotherapeutic drugs on the control group and the experimental group. The results showed that the OD value( absorbance) of cells in the experimental group was significantly lower than that in the control group( P < 0. 05),and the difference was statistically significant. Flow cytometry was used to detect the apoptotic rate of the chemotherapeutic drugs on the control group and the experimental group: apoptosis in the experimental group The expression of MDR gene in the experimental group was significantly lower than that in the QBC939/ADM group( P < 0. 05),and the difference was statistically significant. The expression of MDR gene in the experimental group was significantly lower than that in the QBC939/ADM group( P < 0. 05). Conclusion The resistant cells of human cholangiocarcinoma cell line multidrug QBC939/ADM is successfully established,and preliminary evidence that the Ginsenoside Rg3 of Panax ginseng can effectively reverse the multidrug resistance,but its reversal mechanism has not been studied,should be further explored.