| 王磊,郑广涛.微小RNA-152-3p 对结肠癌细胞增殖、迁移和侵袭的影响及其作用机制研究[J].安徽医药,2022,26(9):1835-1839. |
| 微小RNA-152-3p 对结肠癌细胞增殖、迁移和侵袭的影响及其作用机制研究 |
| Effect of miR-152-3p on proliferation, migration and invasion of colon cancer cells and its mechanism |
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| DOI：10.3969/j.issn.1009-6469.2022.09.032 |
| 中文关键词: 结肠肿瘤 微小RNA-152-3p 转移相关蛋白2 增殖 迁移 侵袭 |
| 英文关键词: Colonic neoplasms miR-152-3p MTA2 Proliferation Migration Invasion |
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| 中文摘要: |
| 目的探讨微小RNA(miR)-152-3p对结肠癌细胞的增殖、迁移和侵袭的影响及机制。方法2019年2月至2020年4月，将结肠癌SW480细胞分为miR-152-3p模拟物阴性对照(miR-NC)组、miR-152-3p模拟物(miR-152-3p)组、抑制物(anti-miRNC)组、miR-152-3p抑制物(anti-miR-152-3p)组、阴性对照(si-NC)组、沉默转移相关蛋白2(si-MTA2)组、miR-152-3p模拟物+空载体(miR-152-3p+pcDNA3.1)组、miR-152-3p模拟物+过表达MTA2(miR-152-3p+pcDNA3.1-MTA2)组，均用脂质体法转染至结直肠癌SW480细胞中，另取未转染结直肠癌SW480细胞记为Control组。采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测miR-152-3p和MTA2 mRNA的表达水平；蛋白质印迹法测定蛋白的表达；MTT法检测细胞活性；Transwell检测细胞迁移和侵袭；双荧光素酶报告基因检测实验检测荧光活性。结果结肠癌HCT116、SW480及LoVo细胞中miR-152-3p表达分别为0.72±0.04、0.29±0.02、0.49±0.01，人正常结肠上皮细胞NCM460中miR-152-3p表达为1.06±0.12，与正常结肠上皮细胞NCM460相比，结肠癌细胞HCT116、LoVo、SW480中MTA2 mRNA和蛋白较高，miR-152-3p较低(P<0.05)；Control组、miR-NC组、miR-152-3p组、si-NC组及si-MTA2组结肠癌SW480细胞吸光度分别为0.92±0.22、0.96±0.17、0.31±0.07、0.99±0.17及0.32±0.09，细胞迁移数分别为(172.00±23.52)个、(169.00±20.66)个、(53.67±12.22)个、(155.67±16.8)个及(54.33±8.74)个，细胞侵袭数分别为(124.67±10.02)个、(122.33±9.45)个、(26.00±5.00)个、(108.33±10.02)个及(42.00±4.00)个，与miR-NC 组相比，miR-152-3p 组SW480细胞中细胞周期蛋白D1(cyclin D1)、基质金属蛋白酶(MMP)-2及SW480细胞活性、迁移和侵袭数量均较低，周期素依赖激酶抑制剂p21(P21)及上皮钙黏素(E-cadherin)较高(P<0.05)；与si-NC组相比，si-MTA2组SW480细胞中cyclin D1、MMP-2及SW480细胞活性、迁移和侵袭数量均较低，P21及E-cadherin较高(P<0.05)。转染miR-152-3p和MTA2野生型表达载体的结肠癌SW480细胞荧光素酶活性显著降低(P<0.05)。相比miR-152-3p+pcDNA3.1组，miR-152-3p+pcDNA3.1-MTA2组SW480细胞中P21、E-cadherin的表达较低，MTA2、cyclin D1、MMP-2蛋白的表达较高，SW480细胞活性、迁移、侵袭数量较高(P<0.05)。结论miR-152-3p可能通过下调MTA2抑制结肠癌细胞的增殖、迁移和侵袭。 |
| 英文摘要: |
| Objective To investigate the effect of miR-152-3p on the proliferation, migration and invasion of colon cancer cells and its mechanism.Methods February 2019 to April 2020, Colon cancer SW480 cells were assigned into miR-152-3p mimic negative control (miR-NC) group, miR-152-3p mimic (miR-152-3p) group, inhibitor (anti-miR-NC) group, miR-152-3p inhibitor (anti-miR-152-3p)group, negative control (si-NC) group, metastasis-associated protein 2 interference (si-MTA2) group, miR -152-3p mimic+empty vector (miR-152-3p+pcDNA3.1) group and miR-152-3p mimic+MTA2 overexpression (miR-152-3p+pcDNA3.1-MTA2) group, colorectal cancer SW480 cells were transfected by liposome method, and untransfected colorectal cancer SW480 cells were taken as the control group. The expressions of miR-152-3p and MTA2 mRNA were detect by quantitative real-time polymerase chain reaction (qRT-PCR);western blotting was used to detect protein expression; methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay was used to detect cell viability; transwell was used to detect cell migration and invasion; the dual luciferase reporter assay was used to detect fluorescence activity.Results The expression of miR-152-3p in colon cancer HCT116, SW480 and LoVo cells was 0.72±0.04, 0.29±0.02, 0.49±0.01,respectively, and the expression of miR-152-3p in normal colonic epithelial cell NCM460 was 1.06±0.12. Compared with cell NCM460,colon cancer cells HCT116, LoVo, SW480 had higher MTA2 mRNA and protein, and lower miR-152-3p (P<0.05). The OD values of colon cancer SW480 cells in the control group, miR-NC group, miR-152-3p group, si-NC group and si-MTA2 group were 0.92±0.22,0.96±0.17, 0.31±0.07, 0.99±0.17 and 0.32±0.09, respectively, and the number of cell migration was 172.00±23.52, 169.00±20.66,53.67±12.22, 155.67±16.8, and 54.33±8.74, and the number of cell invasions was 124.67±10.02, 122.33±9.45, 26.00±5.00, 108.33±10.02 and 42.00±4.00, compared with miR-NC group, cyclin D1, matrix metalloproteinase (MMP)-2 and the viability, migration and invasion numbers of SW480 cells in miR-152-3p group were decreased, and cyclin-dependent kinase inhibitor p21 (P21) and epithelial cadherin (E-cadherin) were increased (P<0.05). Compared with si-NC group, cyclin D1, MMP-2 and the activity, migration and invasion numbers of SW480 cells in si-MTA2 group were lower, and P21 and E-cadherin were higher (P<0.05). The luciferase activity of colon cancer SW480 cells transfected with miR-152-3p and MTA2 wild-type expression vectors was significantly decreased (P<0.05).Compared with the miR-152-3p+pcDNA3.1 group, the expressions of P21 and E-cadherin in SW480 cells in the miR-152-3p+pcDNA3.1-MTA2 group were lower, and the expressions of MTA2, cyclin D1, MMP-2 proteins, and the number of SW480 cell viability, migration and invasion was higher (P<0.05).Conclusion miR-152-3p may inhibit the proliferation, migration and invasion of colon cancer cells by down-regulating MTA2. |
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