多烯磷脂酰胆碱在裸鼠模型中逆转胃癌细胞奥沙利铂耐药的作用及机制研究

Reversal effect and mechanism of polyene phosphatidylcholine on oxaliplatin resistance of gastric cancer cells in a nude mouse model

目的探讨多烯磷脂酰胆碱(PPC)在裸鼠模型中对人胃腺癌耐奥沙利铂细胞株(SGC-7901/L-OHP)的逆转耐药的作用及其机制。方法用继发性耐药细胞株SGC-7901/L-OHP建立裸鼠皮下移植瘤模型,并随机分为4组,空白对照组、多烯磷脂酰胆碱(PPC)组、奥沙利铂(L-OHP)组及奥沙利铂联合多烯磷脂酰胆碱(L-OHP+PPC)组,连续给药14天。21天后处死裸鼠,剥离肿瘤组织,记录肿瘤体积和抑瘤率,对切除的肿瘤组织分别提取肿瘤组织的RNA和蛋白,采用实时荧光定量PCR(qRT-PCR)及Western blot检测TLR4、Nanog及ABCF2表达水平。结果加药处理后,与空白对照组相比,L-OHP组及L-OHP+PPC组移植瘤细胞的增殖均受到抑制(P <0. 001),两药联合有交互作用(P=0. 008); L-OHP+PPC组的体积抑瘤率高于空白对照组(P <0. 001)及L-OHP组(P <0. 001)。L-OHP+PPC组TLR4/Nanog/ABCF2的mRNA表达较空白对照组低,差异有统计学意义(TLR4:P <0. 001; Nanog:P <0. 001; ABCF2:P <0. 001); L-OHP+PPC组TLR4/Nanog/ABCF2的mRNA表达较L-OHP组低,差异有统计学意义(TLR4:P <0. 001; Nanog:P <0. 001; ABCF2:P <0. 001)。两药联合对TLR4/Nanog/ABCF2的mRNA表达均有交互作用,差异有统计学意义(TLR4:P <0. 001; Nanog:P <0. 001; ABCF2:P <0. 001)。L-OHP+PPC组TLR4/Nanog/ABCF2的蛋白表达较空白对照组低,差异有统计学意义(TLR4:P=0. 006; Nanog:P=0. 017; ABCF2:P=0. 014); L-OHP+PPC组TLR4/Nanog/ABCF2的蛋白表达较L-OHP组低,差异有统计学意义(TLR4:P <0. 001; Nanog:P <0. 001; ABCF2:P <0. 001)。两药联合对TLR4/Nanog/ABCF2的蛋白表达均有交互作用,差异有统计学意义(TLR4:P=0. 014; Nanog:P=0. 01; ABCF2:P=0. 015)。结论多烯磷脂酰胆碱在裸鼠模型中对人胃癌耐奥沙利铂细胞株(SGC-7901/L-OHP)有逆转耐药的作用,可能通过影响TLR4、Nanog、ABCF2表达逆转人胃癌耐药细胞的耐药作用。

Objective The Objective of this study was to investigate the effect of polyene phosphatidylcholine（PPC）on the reversal of resistance to human oxaliplatin-resistant cell lines（SGC-7901/L-OHP）in nude mice and its mechanism.Methods The subcutaneous xenograft model of nude mouse was established with a secondary drug-resistant SGC-7901/L-OHP cell line and randomly divided into four groups：blank control group,oxaliplatin group,PPC group and oxaliplatin combined with PPC group.The mice were administered continuously for 14 days.After 21 days,the nude mice were sacrificed and the tumor tissues were removed.The tumor volume and inhibitory rate of tumor were recorded.The RNA and protein of tumor tissue were extracted from the resected tumor tissue.Real-time quantitative PCR（qRT-PCR）and western blot were used to examine the expression of TLR4,Nanog and ABCF2.Results After treatment,the proliferation of transplanted tumor cells in L-OHP group and L-OHP＋PPC group was inhibited compared with the blank control group,P value was 〈0.001,the two drugs combined interaction,P=0.008;The volume inhibition rate of the L-OHP＋PPC group was higher than that of the blank control group（P〈0.001）and the L-OHP group（P〈0.001）.The mRNA expression of TLR4/Nanog/ABCF2 in L-OHP＋PPC group was significantly different from that in the control group（TLR4：P〈0.001;Nanog：P〈0.001;ABCF2：P〈0.001）;The mRNA expression of TLR4/Nanog/ABCF2 in L-OHP＋PPC group was significantly different from that of L-OHP group（TLR4：P〈0.001;Nanog：P〈0.001;ABCF2：P〈0.001）.The combination of the two drugs had an interaction effect on the mRNA expression of TLR4/Nanog/ABCF2（TLR4：P〈0.001;Nanog：P〈0.001;ABCF2：P〈0.001）.The protein expression of TLR4/Nanog/ABCF2 in L-OHP＋PPC group was significantly different from that in the control group（TLR4：P=0.006;Nanog：P=0.017;ABCF2：P=0.014）;The protein expression of TLR4/Nanog/ABCF2 in L-OHP＋PPC group was significantly different from that of L-OHP group（TLR4：P〈0.001;Nanog：P〈0.001;ABCF2：P〈0.001）.The combination of the two drugs had an interaction effect on the protein expression of TLR4/Nanog/ABCF2,and the difference was statistically significant（TLR4：P=0.014;Nanog：P=0.01;ABCF2：P=0.015）.Conclusion PPC has a reversal resistance to human gastric cancer-resistant oxaliplatin（SGC-7901/L-OHP）cell line in nude mouse,possibly by reducing TLR4,Nanog,ABCF2 at the levels of mRNA and proteins to reverses the drug resistance of human gastric cancer resistant cells.