姜黄素逆转食管癌Eca-109/VCR细胞耐药性的研究

目的探讨姜黄素(Cur)通过p38丝裂原活化蛋白激酶(p38MAPK)信号传导通路对人食管癌耐药Eca-109/VCR细胞耐药的逆转作用。方法实验分为空白组、实验A组、实验B组、联合组和低、中、高剂量对照组。空白组给予不含药物的完全培养基进行培养;实验A组给予含20μmol·L^-1 Cur的完全培养基进行培养;实验B组给予含2μg·mL^-1长春新碱(VCR)的完全培养基进行培养;联合组给予含20μmol·L^-1 Cur和2μg·mL^-1VCR的完全培养基进行培养;低、中、高剂量对照组分别给予含10,20和40μmol·mL^-1 SB203580的完全培养基进行培养。干预24 h后,用蛋白质印迹法检测p38MAPK、磷酸化p38MAPK(p-p38MAPK)、切除修复交叉互补蛋白1(ERCC1)和P糖蛋白(P-gp)的表达情况。结果实验A组、实验B组、联合组和低、高剂量对照组的p38MAPK蛋白灰度值分别为1.94×10^4±2927.88,2.09×10^4±651.32,1.50×10^4±1101.53,1.99×10^4±1520.01和1.95×10^4±1875.74,p-p38MAPK蛋白灰度值分别为1.56×10^4±1829.24,1.78×10^4±1718.27,7049.47±1609.40,1.56×10^4±1178.13和4802.11±1842.59,ERCC1蛋白灰度值分别为1.79×10^4±1180.51,1.98×10^4±1245.39,1.34×10^4±1151.07,1.45×10^4±1273.49和6475.16±1293.79,P-gp蛋白灰度值分别为1.42×10^4±996.42,1.64×10^4±1349.59,9614.54±664.98,1.28×10^4±1164.95和6793.34±911.56。实验A组、实验B组和联合组的上述指标与空白组比较,差异均有统计学意义(均P<0.05);低、高剂量对照组的p-p38MAPK、ERCC1和P-gp蛋白表达量与空白组比较,差异均有统计学意义(均P<0.05);联合组的上述指标与实验B组比较,差异均有统计学意义(均P<0.05)。结论有Cur逆转食管癌细胞多药耐药的作用可能与下调p38MAPK通路相关的p38MAPK、p-p38MAPK、ERCC1和P-gp蛋白表达有关。

Curcumin reverses the drug resistance of esophageal cancer Eca-109/VCR cells

Objective To investigate the role of curcumin(Cur)in reversing multidrug resistance of Eca-109/VCR cells through p38 mitogen-activated protein kinase(p38 MAPK)signaling pathway and its possible mechanism.Methods The experiment was divided into blank group,experimental group A,experimental group B,combined group and control–L,-M,-H groups.The blank group was given drug-free complete medium.Experimental group A was given complete culture medium containing 20μmol·L^-1 Cur for culture.Experimental group B was given complete culture medium containing 2μg·mL^-1 vincristine(VCR)for culture.Combined group was treated with 20μmol·L^-1of Cur and 2μg·mL^-1 VCR for culture.The control-L,-M,-H groups were added with 10,20 and 40μmol·mL^-1 of the pathway inhibitor SB203580 for culture.After intervention 24 h,the expression levels of p38 MAPK,phosphorylation-p38 MAPK(p-p38 MAPK),excision repair cross complementary protein 1(ERCC1)and P-glycoprotein(P-gp)proteins were detected by Western Blot.Results The gray values of p38 MAPK in experimental group A,experimental group B,combined group and control-L,-H groups were1.94×10^4±2927.88,2.09×10^4±651.32,1.50×10^4±1101.53,1.99×10^4±1520.01 and 1.95×10^4±1875.74,the gray values of p-p38 MAPK were 1.56×10^4±1829.24,1.78×10^4±1718.27,7049.47±1609.40,1.56×10^4±1178.13 and 4802.11±1842.59,the gray values of ERCC1 were 1.79×10^4±1180.51,1.98×10^4±1245.39,1.34×10^4±1151.07,1.45×10^4±1273.49 and 6475.16±1293.79,the gray values of P-gp were 1.42×10^4±996.42,1.64×10^4±1349.59,9614.54±664.98,1.28×10^4±1164.95 and6793.34±911.56.Compared with the blank group,the above indexes of experimental group A,group B and combination group were significantly different(all P<0.05).The protein expressions of P-P38 MAPK,ERCC1 and P-gp in the low and high dose control groups were significantly different from those in the blank group(all P<0.05).There were significant differences in the above-mentioned indexes between the experimental group B and combined group(all P<0.05).Conclusion The effect of Cur in reversing the multidrug resistance of esophageal carcinoma cells may be related to the down-regulation of p38 MAPK,p-p38 MAPK,ERCC1 and P-gp protein related to p38 MAPK pathway.