联合使用脱甲基化药物和紫杉醇对肾癌细胞的侵袭能力影响机制研究

摘要

目的：观察联合使用脱氧杂胞苷（ DAC）与紫杉醇（ PTX）对肾患细胞是否存在协同作用，并探究其机制。方法离体培养肾透明细胞癌ACHN细胞株，将120培养皿随机分为空白对照组、DAC组、PTX组、联合组，每组30个。 DAC组：用DAC：0．5、1、2、4、8μmol／L处理3 d；PTX组：用PTX：1、2、4、8、16 nmol／L处理3 d；联合组：用DAC＋PTX处理3 d。使用高速分选流式细胞仪观察T肽协同树突状细胞对肾癌细胞的凋亡率；通过ELISA检测淋巴细胞增强因子（LEF1）和E－cadherin在肾癌细胞中的表达；通过ELISA技术检测凋亡蛋白caspass－3在肾癌细胞中的表达。结果 DAC组、PTX组以及联合组对肿瘤细胞的抑制作用明显，其中联合组对肿瘤细胞的抑制作用最强（ P＜0．05），而且DAC和PTX对肿瘤细胞的抑制作用与药物浓度成正比（P＜0．05）。与DAC组、PTX组比较，联合组中LEF1的表达水平最低（P＜0．05），E－cadherin、caspass－3的表达水平最高（P＜0．05），而且DAC和PTX对LEF1表达的抑制作用、对E－cadherin、caspass－3的表达的促进作用与药物浓度成正比（P＜0．05）。结论 DAC与 PTX联合干预肾癌细胞，具有协同作用，而且LEF1是新的肾癌治疗靶点，DAC与 PTX合用可以提高肾癌细胞的凋亡率，并且降低肾癌细胞的侵袭转移能力。%Objective To observe whether combination application of deoxidation cytidine ( DAC) with paclitaxel ( PTX) has a synergistic effect on renal carcinoma cells , and to explore their action mechanism .Methods The renal clear cell carcinoma-ACHN cell line was cultured in vitro .The 120 culture dishes were randomly divided into 4 groups:blank control group,DAC group,PTX group,combination group (DAC+PTX),with 30 dishes in each group.The cells in DAC group were treated by DAC 0.5, 1, 2, 4, 8μmol/L, respectively for 3 days;the celles in PTX group were treated by PTX 1, 2, 4, 8, 16 nmol/L, respectively for 3 days; the cells in combination group were treated by DAC +PTX for 3 days.The apoptosis rates of renal carcinoma cells induced by T peptide combined with dendritic cells were detected by flow cytometry , and the expression levels of lymphocyte enhancement factor (LEF1),E-cadherin and Caspass-3 in renal carcinoma cells were detected by ELISA .Results The inhibitory effects on tumor cells in DAC group , PTX group and combination group were obvious,in which the inhibitory effects in combination group were the most obvious ( P <0.05).Moreover the inhibitory effects on tumor cells in DAC group and PTX group were in direct proportion to the drug concentrations ( P <0.05).As compared with those in DAC group and in PTX group , the expression levels of LEF 1 in combination group were the lowest (P<0.05), however, the expression levels of E-cadherin and Caspass-3 were the highest.Moreover the inhibitory effects of DAC and PTX on the expressions of LEF 1 as well as the promotive effects on the expressions of E-cadherin ,Caspass-3 were in opposite proportion to the drug concentrations ( P <0.05).Conclusion The combination intervention of DAC and PTX on renal carcinoma cells has a synergistic effect .Moreover LEF1 is an new therapeutic target for renal carcinoma .The combination application of DAC with PTX can increase the apoptosis rate of renal carcinoma cells , and can decrease the abilities of invasion and metastasis of renal carcinoma cells .