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miR-3200-3p通过靶向抑制RNF111促进结直肠癌细胞恶性进展的机制研究
引用本文:武 月,殷红专.miR-3200-3p通过靶向抑制RNF111促进结直肠癌细胞恶性进展的机制研究[J].现代肿瘤医学,2022,0(19):3459-3466.
作者姓名:武 月  殷红专
作者单位:1.中国医科大学附属盛京医院急诊科;2.结直肠肛门外科,辽宁 沈阳 110000
基金项目:辽宁省自然科学基金(编号:201602869)
摘    要:目的:探讨miR-3200-3p能否通过靶向抑制RNF111促进结直肠癌细胞恶性进展。方法:选择5株人结直肠癌细胞系,Real-time PCR检测miR-3200-3p、RNF111的表达,Western blot检测RNF111蛋白的表达;利用双荧光素酶实验验证miR-3200-3p与RNF111的靶向抑制;利用miR-3200-3p mimic/inhibitor或利用miR-3200-3p inhibitor与RNF111 siRNA共转染HCT116细胞,Real-time PCR检测miR-3200-3p表达,Western blot检测RNF111、p-SMAD2蛋白表达,MTT检测细胞增殖,划痕实验检测细胞迁移,Transwell检测细胞侵袭,流式细胞术检测细胞凋亡情况。结果:五株人结直肠癌细胞中,HCT116细胞中miR-3200-3p相对高表达,RNF111相对低表达;miR-3200-3p mimic或inhibitor转染后,与各自NC组相比,可显著促进或抑制RNF111蛋白的表达,促进或抑制细胞的增殖、侵袭及迁移,抑制或促进细胞凋亡(均P<0.05);与pmirGLO-MUT 3' UTR+mimics转染组相比,pmirGLO-WT 3' UTR+mimics转染组荧光素酶活性显著降低(P<0.05);与miR-3200-3p NC组相比,miR-3200-3p inhibitor及miR-3200-3p inhibitor+siRNA NC组RNF111蛋白表达显著升高,p-SMAD2蛋白表达显著降低,细胞增殖、迁移及侵袭被显著抑制,细胞凋亡被显著促进(均P<0.05),与miR-3200-3p inhibitor+siRNA NC组相比,miR-3200-3p inhibitor+RNF111 siRNA组细胞RNF111、p-SMAD2蛋白表达、增殖、迁移、侵袭及凋亡被显著逆转(均P<0.05)。结论:miR-3200-3p可通过靶向抑制RNF111促进结直肠癌细胞增殖、侵袭及迁移,抑制细胞凋亡。

关 键 词:结直肠癌  miR-3200-3p  RNF111  增殖  侵袭  迁移  凋亡

Study on the mechanism of miR-3200-3p promoting the malignant progression of colorectal cancer cells through targeted inhibition of RNF111
WU Yue,YIN Hongzhuan.Study on the mechanism of miR-3200-3p promoting the malignant progression of colorectal cancer cells through targeted inhibition of RNF111[J].Journal of Modern Oncology,2022,0(19):3459-3466.
Authors:WU Yue  YIN Hongzhuan
Affiliation:1.Emergency Department;2.Department of Colorectal and Anal Surgery,Shengjing Hospital,China Medical University,Liaoning Shenyang 110000,China.
Abstract:Objective:To explore whether miR-3200-3p can promote the malignant progression of colorectal cancer cells by targeting to inhibit RNF111.Methods:Five human colorectal cancer cell lines were selected.The expression of miR-3200-3p and RNF111 was detected by Real-time PCR,and the expression of RNF111 protein was detected by Western blot.The targeted inhibition of miR-3200-3p and RNF111 was verified by double luciferase experiment.HCT116 cells were transfected with miR-3200-3p mimic/inhibitor or co-transfected with miR-3200-3p inhibitor and RNF111 siRNA.The expression of miR-3200-3p was detected by Real-time PCR,the expression of RNF111 and p-SMAD2 protein was detected by Western blot,cell proliferation was detected by MTT,cell migration was detected by scratches,cell invasion was detected by Transwell,and cell apoptosis was detected by flow cytometry.Results:In five human colorectal cancer cells,miR-3200-3p was relatively high in HCT116 cells,while RNF111 was relatively low.Compared with NC group,miR-3200-3p mimic or inhibitor transfection can significantly promote or inhibit the expression of RNF111 protein,promote or inhibit cell proliferation,invasion and migration,and inhibit or promote cell apoptosis (all P<0.05).Compared with the pmirGLO-MUT 3' UTR+mimics transfection group,the luciferase activity of pmirGLO-WT 3' UTR+mimics transfection group decreased significantly (P<0.05).Compared with miR-3200-3p NC group,the expression of RNF111 protein in miR-3200-3p inhibitor and miR-3200-3p inhibitor+siRNA NC group was significantly increased,p-SMAD2 protein was significantly decreased,cell proliferation,migration and invasion were significantly inhibited,and cell apoptosis was significantly promoted (P<0.05).Compared with miR-3200-3p inhibitor+siRNA NC group,the expression of RNF111 protein and p-SMAD2 protein,proliferation,migration,invasion and apoptosis in the miR-3200-3p inhibitor+RNF111 siRNA group were significantly reversed (P<0.05).Conclusion:miR-3200-3p can promote the proliferation,invasion and migration of colorectal cancer cells and inhibit cell apoptosis by targeted inhibition of RNF111.
Keywords:colorectal cancer  miR-3200-3p  RNF111  proliferation  invasion  migration  apoptosis
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