«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《现代肿瘤医学》[ISSN:1672-4992/CN:61-1415/R]

期数:

2022年19期

页码:

3459-3466

栏目:

论著（基础研究）

出版日期:

2022-08-30

文章信息/Info

Title:

Study on the mechanism of miR-3200-3p promoting the malignant progression of colorectal cancer cells through targeted inhibition of RNF111

作者:

武 月¹; 殷红专²

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

Author(s):

WU Yue¹; YIN Hongzhuan²

1.Emergency Department;2.Department of Colorectal and Anal Surgery,Shengjing Hospital,China Medical University,Liaoning Shenyang 110000,China.

关键词:

结直肠癌; miR-3200-3p; RNF111; 增殖; 侵袭; 迁移; 凋亡

Keywords:

colorectal cancer; miR-3200-3p; RNF111; proliferation; invasion; migration; apoptosis

分类号:

R735.3

DOI:

10.3969/j.issn.1672-4992.2022.19.003

文献标识码:

A

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。

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.

参考文献/References

［1］LA VECCHIA S,SEBASTIAN C.Metabolic pathways regulating colorectal cancer initiation and progression［J］.Semin Cell Dev Biol,2020,98:63-70.
［2］LIU Y,DEGUCHI Y,TIAN R,et al.Pleiotropic effects of PPARD accelerate colorectal tumorigenesis,progression,and invasion［J］.Cancer Res,2019,79(5):954-969.
［3］AIZAWA T,KARASAWA H,FUNAYAMA R,et al.Cancer-associated fibroblasts secrete Wnt2 to promote cancer progression in colorectal cancer［J］.Cancer Med,2019,8(14):6370-6382.
［4］WANG J,CHU XQ,ZHANG D,et al.Knockdown of long non-coding RNA PEG10 inhibits growth,migration and invasion of gastric carcinoma cells via up-regulating miR-3200［J］.Neoplasma,2018,65(5):769-778.
［5］ZHOU F,WANG W,XING Y,et al.NF-kappaB target microRNAs and their target genes in TNFalpha-stimulated HeLa cells［J］.Biochim Biophys Acta,2014,1839(4):344-354.
［6］CHO O,KIM DW,CHEONG JY.Plasma exosomal miRNA levels after radiotherapy are associated with early progression and metastasis of cervical cancer:A pilot study［J］.J Clin Med,2021,10(10):2110.
［7］SHARMA V,ANTONACOPOULOU AG,TANAKA S,et al.Enhancement of TGF-beta signaling responses by the E3 ubiquitin ligase Arkadia provides tumor suppression in colorectal cancer［J］.Cancer Res,2011,71(20):6438-6449.
［8］PIDIKOVA P,REIS R,HERICHOVA I.miRNA clusters with down-regulated expression in human colorectal cancer and their regulation［J］.Int J Mol Sci,2020,21(13):4633.
［9］LIU G,LI B.Role of miRNA in transformation from normal tissue to colorectal adenoma and cancer［J］.J Cancer Res Ther,2019,15(2):278-285.
［10］CIRILLO F,CATELLANI C,SARTORI C,et al.Obesity,insulin resistance,and colorectal cancer:Could miRNA dysregulation play a role［J］.Int J Mol Sci,2019,20(12):2922.
［11］PAN M,CHEN Q,LU Y,et al.MiR-106b-5p regulates the migration and invasion of colorectal cancer cells by targeting FAT4［J］.Biosci Rep,2020,40(11):BSR20200098.
［12］LI C,DING D,GAO Y,et al.MicroRNA3651 promotes colorectal cancer cell proliferation through directly repressing Tbox transcription factor 1［J］.Int J Mol Med,2020,45(3):956-966.
［13］CHEN E,LI Q,WANG H,et al.MiR-92a promotes tumorigenesis of colorectal cancer,a transcriptomic and functional based study［J］.Biomed Pharmacother,2018,106:1370-1377.
［14］LIU W,RUI H,WANG J,et al.Axin is a scaffold protein in TGF-beta signaling that promotes degradation of Smad7 by Arkadia［J］.EMBO J,2006,25(8):1646-1658.
［15］MIYAZONO K,KOINUMA D.Arkadia-beyond the TGF-beta pathway［J］.J Biochem,2011,149(1):1-3.
［16］CHEN H,YANG T,LWI Z,et al.RNF111/Arkadia is regulated by DNA methylation and affects TGF-beta/Smad signaling associated invasion in NSCLC cells［J］.Lung Cancer,2015,90(1):32-40.
［17］HEDRICK E,MOHANKUMAR K,SAFE S.TGFbeta-induced lung cancer cell migration is NR4A1-dependent［J］.Mol Cancer Res,2018,16(12):1991-2002.
［18］MIZUTANI A,KOINUMA D,SEIMIYA H,et al.The Arkadia-ESRP2 axis suppresses tumor progression:analyses in clear-cell renal cell carcinoma［J］.Oncogene,2016,35(27):3514-3523.
［19］KOSTEK O,HACIOGLU MB,SAKIN A,et al.Regorafenib or rechallenge chemotherapy:which is more effective in the third-line treatment of metastatic colorectal cancer［J］.Cancer Chemother Pharmacol,2019,83(1):115-122.
［20］BEKE-SOMFAI T,LINCOLN P,NORDEN B.Double-lock ratchet mechanism revealing the role of alphaSER-344 in FoF1 ATP synthase［J］.Proc Natl Acad Sci USA,2011,108(12):4828-4833.
［21］PETRI BJ,KLINGE CM.Regulation of breast cancer metastasis signaling by miRNAs［J］.Cancer Metastasis Rev,2020,39(3):837-886.

备注/Memo

备注/Memo:

辽宁省自然科学基金（编号：201602869）

常用功能

更新日期/Last Update: 2022-08-31