摘要
目的:观察转染miR-193a-3p结肠癌细胞SW480增殖、迁移和侵袭能力变化及过表达神经前体细胞表达下调蛋白9(NEDD9)对其影响。方法:将购自美国菌种保藏中心结肠癌细胞SW480分为NC组、miR-con组、miR-193a-3p组、si-co组、si-NEDD9组、miR-193a-3p+pcDNA组、miR-193a-3p+pcDNA-NEDD9组,每组9例。采用蛋白质印迹法(Western Blot)检测NEDD9、细胞周期蛋白D1(Cyclin D1)、细胞核增殖抗原标记物(Ki67)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、神经钙黏蛋白(N-cadherin)、上皮钙黏蛋白(E-cadherin)和波形蛋白(Vimentin)表达水平;采用四甲基偶氮唑盐比色法(MTT)检测细胞存活率;采用Transwell检测细胞迁移和侵袭数量。结果:转染miR-193a-3p后,miR-193a-3p组结肠癌细胞SW480存活率、迁移和侵袭数量及相关蛋白Cyclin D1、Ki-67、MMP-2、MMP-9表达水平明显低于NC组和miR-con组(均P<0.01)。沉默NEDD9时,si-NEDD9组上述指标检测水平显著低于NC组和si-con组(均P<0.01)。过表达NEDD9时,上述指标检测水平显著高于miR-193a-3p+pcDNA组(P<0.01)。过表达miR-193a-3p,E-cadherin表达水平较miR-con组升高,N-cadherin、Vimentin表达水平降低(P<0.01)。结论:转染miR-193a-3p可抑制结直肠癌细胞的增殖、迁移和侵袭,过表达NEDD9可能逆转这种作用。
Objective:To observe the effects of miR-193a-3p on colon cancer cell line SW480 proliferation,migration and invasion,and overexpression of neural precursor cells expressed developmentally down-regulated 9(NEDD9).Method:The colon cancer cells SW480 purchased from the American type culture collection were divided into NC group,miR-con group,miR-193a-3p group,si-con group,si-NEDD9 group,miR-193a-3p+pcDNA group,miR-193a-3p+pcDNA-NEDD9 group,each group was 9 cases.Western blot was used to detect NEDD9,Cyclin D1,nuclear proliferation antigen markers(Ki67),matrix metalloproteinase 2(MMP-2),matrix metalloproteinase-9(MMP-9),neuro cadherin(N-cadherin),epithelial cadherin(E-cadherin)and Vimentin protein expression;tetramethylazozolium salt colorimetry(MTT)was used to detect cell viability;Transwell was used to detect cell number of migration and invasion.Results:After transfection with miR-193a-3p,the survival rate,migration and invasion of colon cancer cells SW480 and the expressions of related proteins CyclinD1,Ki-67,MMP-2,MMP-9 in miR-193a-3p group were lower than those in NC group and miR-con group(P<0.05).When NEDD9 was silenced,the detection level of the above indicators in the si-NEDD9 group was significantly lower than that in the NC group and the si-con group(P<0.05).When NEDD9 was overexpressed at the same time,the detection level of the above indicators was significantly higher than that of miR-193a-3p+pcDNA group(P<0.05).Overexpression of miR-193a-3p,the expression of E-cadherin was higher than that of miR-con group,and the expressions of N-cadherin and Vimentin were decreased(P<0.05).Conclusion:Transfection of miR-193a-3p can inhibit the proliferation,migration and invasion of colorectal cancer cells,and overexpression of NEDD9 may reverse this effect.
作者
黄裕文
高昆
任显坤
HUANG Yu-wen;GAO Kun;REN Xian-kun(Deportment of General Surgery,Qionglai Medical Center Hospital,Sichuan Province,Qionglai 611500,China;Department of Emergency surgery,Qionglai Medical Center Hospital,Sichuan Province,Qionglai 611500,China;Department of Gastrointestinal Surgery,Qionglai Medical Center Hospital,Sichuan Province,Qionglai 611500,China)
出处
《微循环学杂志》
2020年第3期19-24,共6页
Chinese Journal of Microcirculation
