摘要
目的:探讨微小RNA(miRNA,miR)-205靶向调控单羧酸转运蛋白1(MCT1)对结直肠癌细胞乳酸水平和增殖能力的影响。方法:选取2017年6月至2019年6月新乡市中心医院收集的280例结直肠癌和对应癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)和蛋白质印迹法(Western blot)分析miR-205和MCT1表达水平;采用慢病毒介导对照miRNA和miR-205感染SW480结直肠癌细胞系,构建miRNA对照组和miR-205组细胞系,采用细胞计数试剂盒(CCK-8)分析两组细胞增殖能力;采用比色法测定肿瘤组织和两组细胞胞内乳酸水平;采用碘化丙锭测定两组细胞周期;采用双荧光素酶报告基因分析miR-205靶基因;采用Western blot分析两组细胞增殖和周期相关蛋白的表达水平。应用SPSS 17.0统计学软件分析,符合正态分布的以均数±标准差表示,组间比较采用t检验。结果:癌旁组织miR-205表达水平(1.22±0.19)明显高于结直肠癌组织(0.51±0.11),差异有统计学意义(t=4.991,P<0.05)。癌旁组织MCT1蛋白相对表达水平(0.87±0.17)明显低于结直肠癌组织(2.72±0.23),差异有统计学意义(t=5.198,P<0.05)。癌旁组织乳酸水平[(1.97±0.24)mmol/L]明显低于结直肠癌组织[(4.71±0.87)mmol/L],差异有统计学意义(t=3.718,P<0.05)。miRNA对照组细胞乳酸水平[(2.76±0.82)mmol/L]明显高于miR-205组[(1.20±0.18)mmol/L],差异有统计学意义(t=3.338,P<0.05)。miRNA对照组细胞吸光度值[(1.86±0.64)]明显高于miR-205组[(1.23±0.39)],差异有统计学意义(t=3.011,P<0.05)。miRNA对照组细胞G 0+G 1期比例[(32.73±3.09)%]明显低于miR-205组[(43.19±3.12)%],差异有统计学意义(t=2.810,P<0.05)。miRNA对照组细胞S期比例[(34.22±4.17)%]明显高于miR-205组[(24.44±2.95)%],差异有统计学意义(t=2.422,P<0.05)。生物信息学分析结果显示MCT1是miR-205的靶基因。miRNA对照组细胞MCT1相对表达水平(1.09±0.12)明显高于miR-205组(0.48±0.10),差异有统计学意义(t=2.864,P<0.05)。结论:miR-205通过调控MCT1表达水平,调节细胞内乳酸水平,进而促进了肿瘤细胞增殖。
Objective To investigate the effects of microRNA(miRNA,miR)-205 on lactate level and proliferation of colorectal cancer cells by targeting monocarboxylate transporter 1(MCT1).Methods Totally,280 cases of colorectal cancer and corresponding adjacent tissues collected in our hospital from June 2017 to June 2019 were selected as the research objects.The expression levels of miR-205 and MCT1 were analyzed by fluorescent quantitative polymerase chain reaction(PCR)and Western blotting.SW480 colorectal cancer cell lines were infected with lentivirus mediated control miRNA and miR-205 to construct miRNA control group and miR-205 group.Cell proliferation was analyzed by cell counting kit-8(CCK-8).The levels of lactate in tumor tissues and cells were measured by colorimetry.The cell cycle was measured by propidium iodide.The miR-205 target gene was analyzed by dual luciferase reporter gene.The targeting protein expression levels were analyzed by Western blotting.SPSS 17.0 statistical software was used to analyze the data.The data in accordance with normal distribution were expressed as mean±standard deviation.Results The expression level of miR-205 in adjacent tissues(1.22±0.19)was significantly higher than that in colorectal cancer tissues(0.51±0.11,t=4.991,P<0.05).The relative expression level of MCT1 protein in adjacent tissues(0.87±0.17)was significantly lower than that in colorectal cancer tissues(2.72±0.23,t=5.198,P<0.05).The lactate level of adjacent tissues[(1.97±0.24)mmol/L]was significantly lower than that of colorectal cancer[(4.71±0.87)mmol/L,t=3.718,P<0.05].The lactate level in miRNA control group[(2.76±0.82)mmol/L]was significantly higher than that in miR-205 group[(1.20±0.18)mmol/L,t=3.338,P<0.05].The absorbance value in miRNA control group[(1.86±0.64)]was significantly higher than that in miR-205 group[(1.23±0.39),t=3.011,P<0.05].The proportion of cells in G0+G1 phase in miRNA control group[(32.73±3.09)%]was significantly lower than that in miR-205 group[(43.19±3.12)%,t=2.810,P<0.05].The proportion of cells in S phase in miRNA control group[(34.22±4.17)%]was significantly higher than that in miR-205 group[(24.44±2.95)%,t=2.422,P<0.05].Bioinformatics showed that MCT1 was the target gene of miR-205.The relative expression level of MCT1 in miRNA control group(1.09±0.12)was significantly higher than that in miR-205 group(0.48±0.10,t=2.864,P<0.05).Conclusion MiR-205 can promote the proliferation of tumor cells by regulating the expression level of MCT1 and the level of intracellular lactic acid.
作者
李智永
杜太平
陈颢
尚卿
李建华
韩玲
刘得静
Li Zhiyong;Du Taiping;Chen Hao;Shang Qing;Li Jianhua;Han Ling;Liu Dejing(Department of General Surgery,Xinxiang Central Hospital,the Fourth Clinical College of Xinxiang Medical College,Xinxiang 453000,China;Department of Neurosurgery,the First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453000,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第6期1086-1089,共4页
Chinese Journal of Experimental Surgery
关键词
微小RNA
结直肠癌
增殖
周期
MicroRNA
Colorectal cancer
Proliferation
Cycle
