多烯磷脂酰胆碱对胃癌奥沙利铂耐细胞株BGC823/L-OHP耐药逆转作用的研究

Effect of polyene phosphatidylcholine on reversing drug-resistance in human oxaliplatin-resistant gastric cancer cell line BGC823/L-OHP

目的探讨多烯磷脂酰胆碱(PPC)对胃癌耐奥沙利铂(L-OHP)细胞株BGC823/L-OHP的增殖抑制、逆转耐药作用及可能机制。方法采用CCK-8法检测不同浓度PPC(0、0.5、1.5、3、6、12、24、48、64、86μmol/L)处理BGC823/L-OHP细胞的增殖率,并检测不同浓度PPC(1、5、10μmol/L)联合不同浓度L-OHP(1、5、10、20、30、40μg/ml)处理BGC823细胞和BGC823/L-OHP细胞48 h的增殖抑制率,检测L-OHP的半数抑制浓度(IC_(50))和逆转耐药作用,同时选取IC_(50)最小时PPC浓度作为后续实验浓度。实时荧光定量PCR(QPCR)和Western blotting检测Toll样受体-4(TLR-4)、Nanog和ABCF2的mRNA和蛋白表达情况。结果 0.5~48μmol/L PPC能促进BGC823/L-OHP细胞增殖,64、86μmol/L PPC可抑制BGC823/L-OHP细胞增殖。经PPC处理,L-OHP作用48 h的IC_(50)由28.54μg/ml下降至6.63μg/ml,逆转倍数为4.31。5μmol/L PPC能显著增加L-OHP对BGC823/L-OHP细胞的敏感性,选取该浓度用于后续实验。与BGC823组比较,BGC823/L-OHP组中ABCF2、TLR-4和Nanog的mRNA和蛋白水平显著上调(P<0.05);经5μmol/L PPC处理后,BGC823/L-OHP组中ABCF2、TLR-4和Nanog的mRNA和蛋白水平显著下调(P<0.05)。结论 PPC联合L-OHP能够增加BGC823/L-OHP细胞株对L-OHP的敏感性,抑制细胞生长,从而逆转耐药。其机制可能是通过影响TLR-4、Nanog和ABCF2表达来发挥逆转耐药的作用。

Objective To investigate the proliferative inhibitory effect and reversal of oxaliplatin（ L-OHP） resistance of polyene phosphatidylcholine（ PPC） on gastric cancer BGC823/L-OHP cells and its possible mechanism. Methods CCK-8 assay was used to detect the effect of PPC（ 0,0. 5,1. 5,3,6,12,24,48,64,86 μmol/L） on the proliferation of BGC823/L-OHP cells. Meanwhile,the effect of PPC（ 1,5,10 μmol/L） in combination with L-OHP（ 1,5,10,20,30,40 μg/ml） on the proliferation of BGC823 cells and BGC823/L-OHP cells at 48 h was detected by CCK-8 assay. The effect of reversing drug resistance and the half maximal inhibitory concentration（ IC50） of L-OHP were determined. The minimum IC50 of PPC was chosen for the following experiment.The expressions of Toll like receptor-4（ TLR-4）,Nanog and ABCF2 mRNA and protein were detected by QPCR and Western blotting.Results Low concentration of PPC（ 0. 5-48 μmol/L） could promote the proliferation of BGC823/L-OHP cells, but high concentration of PPC（ 64,86 μmol/L） could inhibit the proliferation of BGC823/L-OHP cells. The reversal effect of 5 μmol/L PPC was the most obvious. With the treatment of PPC,the IC50 of L-OHP on BGC823/L-OHP cells decreased from 28. 54 μg/ml to 6. 63μg/ml,and the reverse index was 4. 31. The concentration of 5 μmol/L PPC could significantly increase the sensitivity of BGC823/LOHP cells to L-OHP. Compared with BGC823 group,the expressions of ABCF2,TLR-4 and Nanog mRNA and protein in BGC823/LOHP group were significantly up-regulated（ P〈0. 05）,and ABCF2,TLR-4 and Nanog mRNA and protein levels were significantly down-regulated after 5 μmol/L PPC treatment（ P〈0. 05）. Conclusion PPC in combination with L-OHP can increase the sensitivity of BGC823/L-OHP cells to L-OHP,inhibit the growth of BGC823/L-OHP cells and reverse the drug resistance. Its mechanism may play a role in reversing drug resistance by affecting the expression of TLR-4,Nanog and ABCF2.