敲低长链非编码RNA THOR对结直肠癌细胞糖代谢重编程和转移能力的影响

Effect of lncRNA THOR Knockdown on Glucose Metabolic Reprogramming and Metastasis Ability of Colorectal Cancer Cells

目的探讨长链非编码RNA(long non-coding RNA,lncRNA)THOR对结直肠癌(colorectal cancer,CRC)细胞糖代谢重编程、迁移和侵袭能力的影响,以及其与果糖-1,6-二磷酸醛缩酶C(fructose-1,6-bisphosphate aldolase C,ALDOC)之间的靶向调控关系。方法采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)法检测THOR在CRC细胞系中的表达,使用THOR特异性siRNA转染HCT116和SW480细胞沉默THOR的表达,通过细胞划痕实验和Transwell实验检测THOR对CRC细胞迁移、侵袭能力的影响,通过检测CRC细胞的葡萄糖摄取量及丙酮酸生成量探讨THOR对CRC细胞糖代谢重编程的影响,采用RT-qPCR法和Western blot法检测ALDOC mRNA和蛋白质的表达水平。结果RT-qPCR法检测结果显示,THOR在HCT116和SW480细胞中的表达水平显著高于正常人肠上皮细胞NCM460。沉默THOR后,细胞划痕实验及Transwell实验结果表明,HCT116和SW480细胞的迁移与侵袭能力均受到抑制,葡萄糖摄取量及丙酮酸生成量均下降,RT-qPCR法和Western blot法检测结果显示,ALDOC mRNA及蛋白的表达水平显著下调,差异均有统计学意义(P<0.05)。结论lncRNA THOR在CRC细胞中高表达,可能通过靶向调控ALDOC影响CRC细胞糖代谢重编程促进细胞的迁移与侵袭。

Objective To investigate the effects of long non-coding RNA(lncRNA)THOR on glucose metabolism reprogramming,migration and invasion of colorectal cancer(CRC)cells,and its targeted regulatory relationship with fructose-1,6-bisphosphate aldolase C(ALDOC).Methods Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of THOR in CRC cell lines.THOR specific siRNA was transfected into HCT116 and SW480 cells to silence the expression of THOR,and the effects of THOR on the invasion and migration ability of CRC cells was detected by cell scratch assay and Transwell assay.The effect of THOR on glucose metabolic reprogramming in CRC cells was investigated by measuring the glucose intake and pyruvate production in CRC cells,and the expression levels of ALDOC mRNA and protein were detected by RT-qPCR and western blot.Results The results of RT-qPCR showed that the expression levels of THOR in HCT116 and SW480 cells were significantly higher than those in normal human intestinal epithelial cells NCM460.After the silencing of THOR,the cell scratch assay and Transwell assay showed that the migration and invasion ability of HCT116 and SW480 cells were inhibited,and the glucose uptake and pyruvate yield were decreased.The results of RT-qPCR and western blot showed that the expression levels of ALDOC mRNA and protein were significantly down-regulated,the difference was statistically significant(P<0.05).Conclusion lncRNA THOR is highly expressed in CRC cells,which may affect the glucose metabolism reprogramming by targeting ALDOC to promote the migration and invasion of CRC cells.