二甲双胍诱导结直肠癌细胞发生铜死亡及其机制  

Copper death in colorectal cancer cells induced by metformin and its mechanism

作　　者：梁慧慧 唐科 谭亲友 LIANG Huihui;TANG Ke;TAN Qinyou(College of Pharmacy,Guilin Medical University,Guilin 541199,China;Department of Clinical Pharmacy and Clinical Pharmacology,Affiliated Hospital of Guilin Medical University,Guilin 541001,China;China US Health and Disease Lipid Research Center,Guilin 541001,China)

机构地区：[1]桂林医学院药学院,桂林541199 [2]桂林医学院附属医院临床药学与临床药理学教研室,桂林541001 [3]中美健康与疾病脂质研究中心,桂林541001

出　　处：《华夏医学》2025年第1期28-36,共9页Acta Medicinae Sinica

基　　金：国家自然科学基金地区科学基金项目(82160765);广西自然科学基金面上项目(2018GXNSFAA050147);广西博士研究生创新项目(YCSW2023415)。

摘　　要：目的探讨二甲双胍诱导结直肠癌(CRC)细胞铜死亡作用及其机制。方法体外培养CRC细胞系(SW480、HT29),通过CCK8检测CRC细胞经不同浓度的Met分别干预24、48、72 h后的细胞活力;平板克隆实验检测Met对CRC细胞的增殖能力;Transwell检测Met对CRC细胞的迁移能力;倒置显微镜下分别观察低、中、高剂量的Met对CRC细胞形态的影响;Met联合铜死亡抑制剂对CRC细胞增殖的影响;qRT-PCR以及蛋白免疫质印迹分别从基因水平及蛋白水平上检测Met对CRC细胞中铁氧还蛋白1(FDX1)、二氢脂酰胺S-乙酰转移酶(DLAT)和热休克蛋白70(HSP70)表达影响。结果随着Met浓度的增加,CRC细胞活力呈浓度以及时间依赖性下降,而加入铜死亡抑制剂能部分逆转Met对CRC细胞活力的抑制作用;Met显著抑制CRC细胞的增殖能力和迁移能力且抑制作用呈浓度依赖性,以上结果均具有统计学意义(P<0.05)。高倍镜下观察到CRC细胞形态发生明显改变;Met在基因和蛋白水平均可显著抑制FDX1与DLAT的表达,促进HSP70的表达(P<0.05)。结论Met可以通过调控FDXI/DLAT通路诱导CRC细胞铜死亡,进而发挥抗CRC的作用。Objective To investigate the effect of metformin(Met)on the copper death and explore the mechanism in colorectal cancer(CRC)cells.Methods CRC cell lines(SW480,HT29)were cultured in vitro,and CCK8 was used to detect the cell viability of CRC cells after treatment with Met for 24,48,72 h.The proliferation ability of CRC cells was detected by plate cloning.The migration ability of CRC cells was detected by Transwell.The effect of low,medium and high dose of Met on the morphology of CRC cells was observed under inverted microscope.The effect of copper death inhibitor on the proliferation of CRC cells induced by Met was observed.qRT-PCR and Western blot were used to detect the gene and protein expressions of ferroredoxin 1(FDX1),dihydrolipoamide S-acetyltransferase(DLAT)and heat shock protein 70(HSP70)in CRC cells,respectively.Results With the increase of Met concentration,the viability of CRC cell was significantly decreased in a concentration-dependent and time-dependent manner.The inhibitory effect of Met on CRC cell viability was partially reversed by adding Cu-death inhibitor.The proliferation and migration ability of CRC cells was significantly inhibited by Met in a concentration-dependent manner,and the results were statistically significant(P<0.05).The morphological changes of CRC cells were observed under high power microscope.The expressionS of FDX1 and DLAT were significantly down-regulated by Met and promote the expression of HSP70 at both gene and protein levels was significantly up-regulated by Met(P<0.05).Conclusion Met induces the copper death in CRC cells by regulating FDXI/DLAT pathway and plays an anti-CRC role.

关 键 词：二甲双胍 结直肠癌 铜死亡 FDX1 DLAT 

分 类 号：R255.3[医药卫生—中医内科学]