circRAD18/miR-516b/PDK1轴调节葡萄糖代谢重编程与结直肠癌增殖的关系  

Relationship between the glucose metabolism reprogramming regulation by the circRAD18/miR-516b/PDK1 axis and proliferation in colorectal cancer cells

作　　者：李威 唐云云[2] 彭娟 蒋训归 LI Wei;TANG Yunyun;PENG Juan;JIANG Xungui(Department of Anorectal Hernia Surgery,Yongzhou Central Hospital,Yongzhou,Hunan 425006,China;Department of Public Fundamentals,Yongzhou Vocational and Technical College,Yongzhou,Hunan 425100,China)

机构地区：[1]湖南省永州市中心医院肛肠疝外科,湖南永州425006 [2]永州职业技术学院公共基础学部,湖南永州425100

出　　处：《中国普通外科杂志》2023年第10期1522-1530,共9页China Journal of General Surgery

摘　　要：背景与目的:环状RNA circRAD18被发现在乳腺癌和甲状腺癌的进展中起了促进作用,但其在其他恶性肿瘤的表达及作用尚未被充分揭示。笔者前期通过生物信息学软件预测circRAD18可与miR-516b互补结合,而葡萄糖代谢关键调节酶丙酮酸脱氢酶激酶1(PDK1)可能是miR-516b的靶基因。因此,本研究初步探讨circRAD18在结直肠癌细胞中的表达及作用,及其对靶miRNA及下游靶基因的调控关系。方法:用qRT-PCR检测不同结直肠癌细胞系(SW480、SW620、HT-29)及正常结直肠上皮细胞(NCM460)中circRAD18的表达;用si-circRAD18沉默结直肠癌细胞中circRAD18的表达后,分别用CCK-8实验和相应的试剂盒检测细胞的增殖情况以及葡萄糖摄取量和乳酸产生量。用双荧光素酶报告基因实验与RNA免疫沉淀(RIP)实验分析circRAD18、miR-516b及PDK1之间的结合关系;最后,采用过表达/敲低实验进一步验证三者之间的关系。结果:与正常结直肠上皮细胞比较,circRAD18在各结直肠癌细胞系中的表达均明显上调(均P<0.05);转染si-circRAD18后,结直肠癌细胞增殖能力、葡萄糖摄取及乳酸产生量均明显降低(均P<0.05);双荧光素酶报告基因实验与RIP实验证实circRAD18可与miR-516b结合,而PDK1是miR-516b的下游靶基因。miR-516b模拟物及si-circRAD18的转染可明显抑制细胞葡萄糖摄取、乳酸产生及PDK1蛋白表达,且补充PDK1可逆转该抑制作用(均P<0.05)。结论:circRAD18在结直肠癌细胞中表达上调,并与结直肠癌细胞增殖能力的增强密切相关,作用机制可能与circRAD18通过海绵样吸附miR-516b后,上调PDK1表达,从而导致结直肠癌细胞葡萄糖代谢重编程有关。Background and Aims:Circular RNA circRAD18 has been found to play a promoting role in the progression of breast cancer and thyroid cancer,but its expression and function in other malignant tumors have not been fully elucidated.In our previous study,we used bioinformatics software to predict that circRAD18 can interact with miR-516b through complementary binding,and pyruvate dehydrogenase kinase 1(PDK1),a key regulator of glucose metabolism,may a target gene of miR-516b.Therefore,this study was conducted to preliminarily investigate the expression and function of circRAD18 in colorectal cancer cells and its regulatory relationship with the target miRNA and downstream target genes.Methods:The expression of circRAD18 in different colorectal cancer cell lines(SW480,SW620,HT-29)and normal colorectal epithelial cells(NCM460)was detected using qRT-PCR.After silencing circRAD18 with si-circRAD18 in colorectal cancer cells,cell proliferation,glucose uptake,and lactate production were assessed using CCK-8 assay and corresponding kits.The binding relationship between circRAD18,miR-516b,and PDK1 was analyzed through dual-luciferase reporter gene assay and RNA immunoprecipitation(RIP)experiment.Finally,overexpression and knockdown experiments were conducted to further validate the relationships among them.Results:Compared to normal colorectal epithelial cells,circRAD18 was significantly upregulated in all colorectal cancer cell lines(all P<0.05).Transfection with si-circRAD18 resulted in a significant decrease in colorectal cancer cell proliferation,glucose uptake,and lactate production(all P<0.05).Dualluciferase reporter gene assay and RIP experiments confirmed the binding of circRAD18 to miR-516b,and PDK1 was identified as a downstream target gene of miR-516b.Transfection with miR-516b mimic or si-circRAD18 significantly inhibited cellular glucose uptake,lactate production,and PDK1 protein expression in colorectal cancer cells,and the supplementation of PDK1 could reverse this inhibitory effect(all P<0.05).Conclusions:CircR

关 键 词：结直肠肿瘤 RNA 环状 代谢 细胞增殖 

分 类 号：R735.3[医药卫生—肿瘤]