姜黄素增强人胆管癌细胞株QBC939对吉西他滨敏感性的研究

Enhanced chemosensitivity of cholangiocarcinoma cell line QBC939 to gemcitabine induced by curcumin

目的探讨姜黄素对人胆管癌细胞株QBC939吉西他滨的增敏作用及其机制研究。方法将人胆管癌细胞QBC939培养后分为对照组、5μg／m1单药姜黄素组、0．625μg／m1单药吉西他滨组及5μg／m1姜黄素联合0．625μg／m1吉西他滨组，采用噻唑蓝（MTT）法检测细胞增殖、流式细胞仪检测细胞周期及凋亡变化，同时用Westernblot法检测相关蛋白核因子±κB（NF±κB）、细胞周期蛋白（Cyclin）B1、凋亡抑制蛋白（1AP）-1的表达改变。结果姜黄素联合吉西他滨处理人胆管癌细胞QBC939后12、24、48、72h的抑制率分别为（10．00±0．02）％、（23．10±0．03）％、（33．234±0．01）％、（48．304±0．05）％，显著高于对照组及单药组（P=0．001、0．000、0．000、0．000）。两药联合作用后能使（28．75±3．23）％的人胆管癌细胞QBC939被阻滞在G2／M期，显著高于单药姜黄素组及单药吉西他滨组[（14．494±2．20）％、（11．824±2．10）％；P=0．000]，且细胞凋亡率为（13．644±3．40）％，显著高于对照组、单药姜黄素组及单药吉西他滨组[（4．534±0．56）％、（6．47±0．98）％、（9．874±1．12）％，P=0．000]；姜黄素联合吉西他滨作用于人胆管癌细胞株QBC939后NF±κB及下游分子CyclinB1和IAP±1蛋白的相对表达量分别为0．3004±0．001、0．4074±0．036、0．4754±0．032，显著低于对照组、单药姜黄素组及单药吉西他滨组（P=0．000、0．001、0．003）。结论姜黄素可增强人胆管癌细胞株QBC939对吉西他滨的敏感性，其机制可能与NF±κB信号通路的抑制相关。

Objective To investigate the effect of curcumin in enhancing chemosensitivity of cholangiocarcinoma cell line QBC939 to gemcitabine and its mechanism. Methods QBC939 cells were divided into control group, 5 μg/mL curcumin group, 0. 625 μg/mL gemcitabine group and 5 trmolYL curcumin combined 0. 625 Ixg/ml gemcitabine group. Methyl thiazol tetrazolium （MTT） assay was used to detect the proliferation inhibition effect, flow cytometry was used to detected the change of cell cycle and apoptosis rate. Expressions of nuclear factor -κB （NF -κB ）, Cyclin B1 and inhibitor of apoptosis proteins （IAP） -1 in different groups were detected by Western blotting. Results Compared with control group, curcumin group and gemcitabine group, the inhibitory rates of cells in curcumin combined gemcitabine group at 12, 24, 48 and 72 h were （10.00 ±0.02）%, （23.10 ±0.03）%, （33.23 ±0.01）%, （48.50 ± 0. 05） % respectively, which were significantly higher than those of control group and single me- dicaments treatment group （P =0. 001,0.000, 0. 000, 0. 000） ; besides, curcumin combined with gem- citabine blocked （28.75 s 3.23 ） % human cholangiocarcinoma cell line QBC939 in G2/M phase, which was significantly higher than those of curcumin group and gemcitabine group [ （14. 49 ±0. 20 ）%, （ 11.82 ±0. 10） % ; P = 0. 000 ], the apoptosis rate in combined group was （ 13.64 ±0 3.40 ） %, signifi- cantly higher than those of control group, curcumin group and gemcitabine group [ （ 4. 53 s 0. 56 ） % , （6. 47 ±0. 98）%, （9. 87 s 1.12）% ; P =0. 000] ; moreover, the relative expression levels of NF -κB and its downstream moleculor Cyclin B1 and IAP - 1 proteins in curcumin combined with gemcitabine group were 0. 300 ±0. 001,0. 407 ±0. 036, 0. 475 ±0. 032 respectively, significantly lower than those of controlgroup, curcumin group and gemcitabine group （P =0.000,0.001,0.003）. Conclusion Curcumin can enhance the sensitivity of human cholangioearcinoma cell line QBC939 to gemcitabine through the inactiva- tion of NF -κB signaling pathway.