卡培他滨通过调控miR-143-5p表达抑制结肠癌细胞生长的研究  

Capecitabine inhibits colon cancer cells growth by regulating the expression of miR-143-5p

作　　者：孙利萍 熊非 肖静 朱顺 SUN Li-ping;XIONG Fei;XIAO Jing;ZHU Shun(Department of Pharmacy,The Eighth Hospital of Wuhan,Wuhan 430010,Hubei Province,China;Department of Anorectal Surgery,The Eighth Hospital of Wuhan,Wuhan 430010,Hubei Province,China;Department of Pharmacy,Wuhan Third Hospital,Wuhan 430062,Hubei Province,China)

机构地区：[1]武汉市第八医院,药学部,湖北武汉430010 [2]武汉市第八医院,肛肠外科,湖北武汉430010 [3]武汉市第三医院药学部,湖北武汉430062

出　　处：《中国临床药理学杂志》2022年第19期2306-2310,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨卡培他滨(CAP)对结肠癌细胞生长的调控作用及其分子作用机制。方法将LoVo细胞分为对照组(正常培养的LoVo细胞)、实验组(使用2 mmol·L^(-1)的卡培他滨处理细胞48 h)、实验组+anti-miR-NC组(转染anti-miR-NC+2 mmol·L^(-1)的卡培他滨)和实验组+anti-miR-143-5p组(转染anti-miR-143-5p+2 mmol·L^(-1)的卡培他滨)。用实时荧光定量聚合酶链反应(RT-PCR)法检测miR-143-5p表达水平,用细胞计数试剂盒8(CCK-8)法检测细胞活力,用流式细胞术检测细胞周期进程和凋亡情况,用蛋白质印迹(Western blot)法检测影响LoVo细胞增殖和凋亡相关蛋白表达水平。结果对照组、实验组、实验组+anti-miR-NC组和实验组+anti-miR-143-5p组的miR-143-5p表达水平分别为1.00±0.14,1.86±0.10,1.81±0.20和1.11±0.13;细胞活力分别为(100.00±2.28)%,(58.05±1.58)%,(50.29±4.44)%和(79.61±6.41)%;G1期细胞比例分别为(53.49±3.00)%,(67.13±3.24)%,(68.82±3.52)%和(56.69±4.31)%;凋亡率分别为(4.14±0.64)%,(21.87±2.89)%,(20.00±2.02)%和(7.75±0.48)%;Ki67蛋白表达水平分别为0.93±0.09,0.42±0.03,0.38±0.05和0.82±0.05;cleaved Caspase-3蛋白表达水平分别为0.34±0.04,0.82±0.07,0.80±0.09和0.43±0.05;磷酸化磷脂酰肌醇3-激酶(p-PI3K)蛋白表达水平分别为0.92±0.07,0.43±0.04,0.37±0.05和0.73±0.10;磷酸化蛋白激酶B(p-AKT)蛋白表达水平分别为0.89±0.11,0.36±0.05,0.39±0.03和0.76±0.05。实验组的上述指标与对照组比较,差异均有统计学意义(均P<0.05);实验组+anti-miR-143-5p组的上述指标与实验组+anti-miR-NC组比较,差异均有统计学意义(均P<0.05)。结论卡培他滨可通过miR-143-5p抑制PI3K/AKT/mTOR信号通路,进而抑制LoVo细胞增殖和周期进程并促进凋亡。Objective To investigate the regulation and molecular mechanism of capecitabine(CAP)on the growth of colon cancer cells.Methods The LoVo cells were divided control group(normal cultured LoVo cells),experimental group(cells treated with 2 mmol·L^(-1) capecitabine for 48 h),experimental+anti-miR-NC(transfected with anti-miR-NC+2 mmol·L^(-1) capecitabine)and experimental+anti-miR-143-5P(transfected with anti-miR-143-5p+2 mmol·L^(-1) capecitabine).Cell viability was detected by cell counting kit-8 method.Real-time polymerase chain reaction was used to detect the relative expression of miR-143-5p.Flow cytometry was used to detect cell cycle progression and apoptosis rate.Western blot was used to detect the protein expression levels of proliferation and apoptosis in LoVo cells.Results The relative expression levels of miR-143-5p in control,experimental,experimental+anti-miR-NC and experimental+anti-miR-143-5p groups were 1.00±0.14,1.86±0.10,1.81±0.20 and 1.11±0.13,respectively;the cell viability were(100.00±2.28)%,(58.05±1.58)%,(50.29±4.44)%and(79.61±6.41)%,respectively;the percentages of G1 phase cells were(53.49±3.00)%,(67.13±3.24)%,(68.82±3.52)%and(56.69±4.31)%,respectively;the apoptosis rates were(4.14±0.64)%,(21.87±2.89)%,(20.00±2.02)%and(7.75±0.48)%,respectively;the Ki67 protein expression were 0.93±0.09,0.42±0.03,0.38±0.05 and 0.82±0.05,respectively;the expression levels of cleaved Caspase-3 were 0.34±0.04,0.82±0.07,0.80±0.09 and 0.43±0.05,respectively;the phospho phosphatidylinositol 3-kinase(p-PI3K)protein expression were 0.92±0.07,0.43±0.04,0.37±0.05 and 0.73±0.10,respectively;the phospho associated protein(p-AKT)protein expression was 0.89±0.11,0.36±0.05,0.39±0.03,0.76±0.05,respectively.Compared the control group with the experimental group,all the above indicators were statistically significant(all P<0.05);Compared the experimental+anti-miR-NC group with the experimental+anti-miR-143-5p group,all the above indicators were statistically significant(all P<0.05).Conclusion Ca

关 键 词：卡培他滨 miR-143-3p 结肠癌 LOVO细胞 凋亡 

分 类 号：R979[医药卫生—药品]