多不饱和脂肪酸对结直肠癌细胞生物学功能的影响及其生物信息学分析

Effect of polyunsaturated fatty acid on biological function of colorectal cancer cells and bioinformatics analysis

目的 探讨ω-3和ω-6多不饱和脂肪酸对结直肠癌细胞的生物学功能影响、联合奥沙利铂对结直肠癌细胞活力的影响,及对结直肠癌细胞基因表达谱的影响。方法 在结直肠癌细胞株HT29和RKO中,三种多不饱和脂肪酸花生四烯酸(AA)、二十二碳六烯酸(DHA)、二十碳五烯酸(EPA)进行干预,CCK-8实验测定细胞活力、流式细胞仪检测细胞凋亡、周期,并与奥沙利铂联合处理检测细胞增殖,AA、EPA处理HT29细胞进行基因芯片的表达谱分析,qRT-PCR对差异基因进行验证分析。结果 三种多不饱和脂肪酸浓度的增加使HT29和RKO细胞的增殖比例显著降低(P<0.05),与奥沙利铂联合后,结直肠癌细胞的活力显著低于单独处理组。AA、DHA、EPA使HT29和RKO的细胞周期阻滞在G0/G1期(P<0.05),凋亡细胞比例与多不饱和脂肪呈浓度依赖关系。AA、EPA干预引起HT29脂肪酸代谢、肿瘤相关等信号通路发生变化,qRT-PCR结果验证结果与芯片结果一致。结论 ω-3和ω-6多不饱和脂肪酸促进细胞凋亡、细胞周期阻滞进而抑制结直肠癌细胞凋亡,增强奥沙利铂的药物效应,作用效果为EPA>DHA>AA;同时肿瘤相关的基因表达谱发生改变。

Objective To investigate the biological effects ofω-3 andω-6 polyunsaturated fatty acids(PUFAs)on colorectal cancer cells,their impact on cell viability when combined with oxaliplatin,and their influence on the gene expression profile of colorectal cancer cells.Methods Colorectal cancer cell lines HT29 and RKO were treated with three types of polyunsaturated fatty acids[arachidonic acid(AA),do cosahexaenoic acid(DHA),and eicosapentaenoic acid(EPA)].Cell viability was assessed using a CCK-8 assay,apoptosis and cell cycle were analyzed using flow cytometry,and cell proliferation was evaluated in combination with oxaliplatin.Gene expression profiling of HT29 cells treated with AA and EPA was conducted using microarrays,and differentially expressed genes were validated by qRT-PCR.Results Increased concentrations of the three polyunsaturated fatty acids significantly reduced the proliferation of HT29 and RKO cells(P<0.05).When combined with oxaliplatin,cell viability was significantly lower compared to individual treatments.AA,DHA,and EPA caused cell cycle arrest at the G0/G1 phase(P<0.05),and the proportion of apoptotic cells showed a concentration-dependent relationship with the polyunsaturated fatty acids.Treatment with AA and EPA led to changes in fatty acid metabolism and tumor-related signaling pathways in HT29 cells.The qRT-PCR results were consistent with the microarray findings.Conclusionω-3 andω-6 polyunsaturated fatty acids promote apoptosis and cell cycle arrest,thereby inhibiting colorectal cancer cell proliferation.They also enhance the drug effects of oxaliplatin,with the effect being the most pronounced for EPA,followed by DHA and then AA.Additionally,the gene expression profile related to tumorigenesis is altered.