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目的研究二十二碳六烯酸(docosahexaenoic acid,DHA)和选择性环氧合酶-2(cyclooxygenase-2,COX-2)抑制剂NS-398联合对人胆管癌细胞QBC939的抑制作用。方法体外培养人胆管癌QBC939细胞株,分别设立DHA浓度组、NS-398浓度组、二者联合组和空白对照组,即将0、15、30、45、60、75μg/ml浓度的DHA和0、25、50、100、150、200μmol/L浓度的NS-398分别联合作用于胆管癌QBC939细胞;应用CCK8法检测其分别对QBC939细胞生长的相对抑制率;采用流式细胞术检测QBC939细胞的凋亡情况;应用Transwell小室检测QBC939细胞的体外侵袭情况。结果 DHA和NS-398在体外均能够单独抑制QBC939细胞生长,45μg/ml的DHA联合100μmol/L的NS-398经24 h作用后,细胞生长抑制率达到90%,实验组与DHA浓度组、NS-398浓度组和空白组差异均有统计学意义(P<0.05),继续增加2种药物浓度,细胞生长抑制率变化不明显;流式细胞术显示DHA和NS-398联合能明显促进QBC939细胞早期凋亡;15μg/ml的DHA联合50μmol/L的NS-398经24 h作用后,对胆管癌细胞生长无明显抑制作用,但QBC939的体外侵袭能力明显降低,与对照组相比差异有统计学意义(P<0.01)。结论 DHA和NS-398联合能明显抑制胆管癌QBC939细胞生长,促进胆管癌细胞早期凋亡,抑制胆管癌细胞的侵袭,二者联合对胆管癌QBC939细胞生长的抑制作用可能是通过促进细胞早期凋亡实现的。
Objective To investigate the inhibitory effect of docosahexaenoic acid (DHA) and selective cyclooxygenase-2 (NSC-2) inhibitor NS-398 on human cholangiocarcinoma cell line QBC939. Methods Human cholangiocarcinoma QBC939 cells were cultured in vitro. DHA concentration and NS-398 concentration groups were established respectively. The combination of the two groups and the blank control group were 0, 15, 30, 45, 60 and 75 μg / ml concentrations of DHA and 0 , 25,50,100,150 and 200μmol / L NS-398, respectively. The relative inhibitory rate of QBC939 cell growth was detected by CCK8 assay. Flow cytometry was used to detect the expression of QBC939 cell Apoptosis was detected in vitro. Transwell chamber was used to detect the in vitro invasion of QBC939 cells. Results DHA and NS-398 could inhibit the growth of QBC939 cells alone in vitro. The growth inhibition rate of DHA and NS-398 was 90% after treated with 45μg / ml DHA and 100μmol / L NS-398 for 24 hours. NS-398 concentration group and the blank group were significantly different (P <0.05), continue to increase the concentration of two drugs, cell growth inhibition rate did not change significantly; flow cytometry showed DHA and NS-398 combination can significantly promote QBC939 The apoptosis of cholangiocarcinoma cells was inhibited by 15μg / ml DHA and 50μmol / L NS-398 for 24 h, but the invasion ability of QBC939 in vitro was significantly lower than that of the control group Statistical significance (P <0.01). Conclusion The combination of DHA and NS-398 can significantly inhibit the growth of human cholangiocarcinoma QBC939 cells, promote the early apoptosis of cholangiocarcinoma cells and inhibit the invasion of cholangiocarcinoma cells. The combination of both could inhibit the growth of QBC939 cholangiocarcinoma cells by promoting early cell apoptosis Realized.