| 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 |
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| 引用本文： | 武 月1,殷红专2. 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. DOI: 10.3969/j.issn.1672-4992.2022.19.003 |
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| 作者姓名： | 武 月1 殷红专2 |
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| 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 |
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| 基金项目： | 辽宁省自然科学基金（编号:201602869） |
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| 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。
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| 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 |
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| WU Yue1,YIN Hongzhuan2. 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. DOI: 10.3969/j.issn.1672-4992.2022.19.003 |
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| Authors: | WU Yue1 YIN Hongzhuan2 |
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| Affiliation: | 1.Emergency Department;2.Department of Colorectal and Anal Surgery,Shengjing Hospital,China Medical University,Liaoning Shenyang 110000,China. |
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| 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. |
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| Keywords: | colorectal cancer miR-3200-3p RNF111 proliferation invasion migration apoptosis |
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