Experimental Study
MicroRNA-29b promotes cellular senescence of colorectal tumors by targeting tribbles pseudokinase 2
Zhenlin Hou, Guihua Wang, Junbo Hu, Jing Wang, Li Sun
Published 2019-12-08
Cite as Chin J Exp Surg, 2019, 36(12): 2203-2206. DOI: 10.3760/cma.j.issn.1001-9030.2019.12.023
Abstract
ObjectiveTo investigate the mechanism of microRNA (miRNA, miR)-29b regulation of tribbles pseudokinase 2 (TRIB2) in colorectal tumors.
MethodsThe online database was used to analyze the miRNAs that bind to the 3’Untranslated Region (UTR) of TRIB2, and the highest scored miR-29b was selected for study. The eukaryotic cell transfection technique was used to interfere with the expression of miR-29b in colorectal tumor cells; Western blotting and real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the difference in protein and mRNA expression of TRIB2; the position of miR-29b binding on TRIB2-3’untranslated regions (3’UTR) was confirmed by luciferase reporter gene assay; β-galactosidase staining was used to detect effects of miR-29b on colorectal cancer cell senescence. All data were quantified as Mean±SD. Two-tailed Student’s t-test was used to evaluate the differences between two groups. All statistical analyses were performed using SPSS 24.0 (SPSS Inc.).
ResultsIn SW48 and SW480 cell lines overexpressing miR-29b, the mRNA levels of TRIB2 decreased to (63.468±4.154)% and (43.145±7.523)% (t=5.351, P<0.01 and t=6.074, P<0.01) compared to the control group. In the cells transfered to the inhibitor targeted miR-29b, the mRNA level of TRIB2 increased to (205.033±27.070)% and (233.353±18.649)% (t=4.663, P<0.01 and t=9.411, P<0.01). Consistent with the above results, miR-29b suppressed, and the inhibitor up-regulated the protein level of TRIB2. After overexpression of miR-29b in colon cancer cells SW48 and SW480, the luciferase activity of the wild-type TRIB2-3’UTR plasmid decreased to (53.531±3.787)% and (45.051±1.728)%, respectively (t=6.928, 15.570, P<0.01), while the luciferase activity of the mutant TRIB2-3’UTR plasmid decreased slightly (decreased to (87.366±3.825)% and (92.839±3.171)%, t=5.578, P<0.01 and t=2.245, P>0.05). In the cancer cells overexpressing of miR-29b, the proportion of senescent cells increased from 41.473% to 62.085% (χ2=19.731, P<0.01), and overexpression of TRIB2 reduced the proportion to 46.866% (χ2=12.031, P<0.01).
ConclusionmiR-29b can inhibits TRIB2 expression and promote the senescence of colorectal tumors, and it can be used as a potential therapeutic target for colorectal cancer.
Key words:
MicroRNA-29b; Tribbles pseudokinase 2; Colorectal cancer; Cell senescence
Contributor Information
Zhenlin Hou
Cancer Research Institute, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China
Colorectal Surgery, Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, China
Guihua Wang
Cancer Research Institute, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China
Junbo Hu
Cancer Research Institute, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China
Jing Wang
Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
Li Sun
Department of Oncology, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China
