结直肠癌中靶向调控SATB2的有关miRNAs的研究  

Analysis of miRNAs related to the target regulation of SATB2 in colorectal cancer based on RT-PCR

作　　者：王军梅[1] 朱李茹[1] 郝世勇[1] WANG Jun-mei;ZHU Li-ru;HAO Shi-yong(Department of Clinical Laboratory,Xiangyang Center Hospital,Xiangyang Hubei 441021,China)

机构地区：[1]襄阳市中心医院检验科

出　　处：《临床和实验医学杂志》2019年第16期1728-1732,共5页Journal of Clinical and Experimental Medicine

摘　　要：目的探讨结直肠癌中靶向调控特异AT序列结合蛋白2(SATB2)有关的miR-31和miR-182的表达水平的意义,并分析其与SATB2的关系。方法从病检确诊为结直肠癌的标本中随机选取6例为实验组,从非癌症的标本中随机选取6例为对照组。利用荧光原位杂交技术和RT-PCR技术,分别检测结直肠癌细胞组织中miRNA的表达情况,以及正常对照组织中候选miRNA的表达情况,分析miRNA和SATB2之间的关系及其临床意义。CCK-8法检测miRNA对体外细胞增值的影响。流式细胞术检测miRNA对细胞周期的影响。裸鼠皮下成瘤比较miRNA对肿瘤增值的影响。结果导入miR-182和miR-31的模拟物后,降低细胞内SATB2表达水平(P<0.05),而将其抑制物导入后,可明显增加SW620和M5细胞中的SATB2表达(P<0.05)。与正常组织相比,结直肠癌组织中miR-182和miR-31表达明显增高(P<0.05)。SW620/miR-31和M5/miR-31细胞增殖能力明显高于对照组细胞的增殖能力(P<0.001)。SW620/miR-182和M5/miR-182细胞增殖能力明显高于对照组细胞的增殖能力(P<0.01)。与对照组相比,miR-31/miR-182组,停留在G1期的细胞减少(P<0.05),S期的细胞增多(P<0.05)。成瘤5 d后,SW620/miR-31和SW620/miR-182与对照组细胞相比,皮下成瘤速度较快(P<0.001)。结论miR-31和miR-182在结直肠癌中高表达与SATB2表达具有显著性相关,同时miR-31和miR-182能反向调控SATB2的表达。Objective To explore the significance of the expression level of miR-31 and miR-182 related to the target regulation of SATB2 in colorectal cancer,and to analyze the relationship with SATB2.Methods Six cases were randomly selected from the colorectal samples diagnosed by pathological diagnosis as the experimental group,and six cases were randomly selected from the non-cancer samples as the control group.The fluorescence in situ hybridization and RT-PCR technique were used to detect the miRNA of colorectal cancer cells in colorectal cancer tissues and normal tissues,and to analyze the relationship between miRNA and SATB2 and its clinical significance.Cck-8 assay was used to detect the effect of miRNA on cell proliferation in vitro.The effect of miRNA on cell cycle was detected by flow cytometry.Effects of miRNA on tumor proliferation were compared in subcutaneous tumorigenesis in nude mice.Results After the introduction of mir-182 and mir-31 mimics,SATB2 expression level in cells was reduced(P<0.05),while SATB2 expression in SW620 and M5 cells was significantly increased after the introduction of its inhibitors(P<0.05).Compared with normal tissues,the expressions of mir-182 and mir-31 in colorectal cancer tissues were significantly higher(P<0.05).The proliferation capacity of SW620/mir-31 and M5/mir-31 cells was significantly higher than that of the control group(P<0.001).The proliferation capacity of SW620/mir-182 and M5/mir-182 cells was significantly higher than that of the control group(P<0.01).Compared with the control group,in the mir-31/mir-182 group,there were fewer cells in G1 phase(P<0.05)and more cells in S phase(P<0.05).After 5 days of tumor formation,the subcutaneous tumor formation rate of SW620/mir-31 and SW620/mir-182 was faster than that of the control cells(P<0.001).Conclusion High expression of miR-31 and miR-182 in colorectal cancer was significantly associated with SATB2 expression,while miR-31 and miR-182 can reverse the expression of SATB2.

关 键 词：结直肠癌 特异AT序列结合蛋白2 miR-31 miR-182 RT-PCR 

分 类 号：R735.34[医药卫生—肿瘤]