Li Yasha,Yang Ke,Zhu Jing,Bi Yang,Tan Bin,Tian Hefeng,Yi Qin,Zhong Haiying.MicroRNA 26b-5p inhibits cell proliferation by targeting p300 in human umbilical cord mesenchymal stem cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(16):1614-1618.
Li Yasha,Yang Ke,Zhu Jing,Bi Yang,Tan Bin,Tian Hefeng,Yi Qin,Zhong Haiying.MicroRNA 26b-5p inhibits cell proliferation by targeting p300 in human umbilical cord mesenchymal stem cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(16):1614-1618.DOI:
Objective To investigate the target microRNA and its effect on the down-regulation of transcriptional coactivator p300
and then on the proliferation of human umbilical cord derived mesenchymal stem cells (hUC-MSCs). Methods Dual luciferase gene reporter system (Firefly/Renilla Luciferase) was used to screen the effective microRNA targets which interacted with p300 3′ UTR. Then the hUC-MSCs were transfected with microRNA mimics
and the expression of p300 3′ UTR at miRNA and protein levels was detected by RT-qPCR and Western blotting respectively. The levels of pluripotency transcription factors including Nanog
Oct4 and Sox2 were measured by RT-qPCR and Western blotting too. MTT assay was employed to detect the proliferation and colony formation in hUC-MSCs before and after the transfection of microRNA mimics. Results It was found that mir 26b-5p binding to p300 3′ UTR with imperfectly matched sequence. The Firefly/Renilla’s relative activity of mir 26b-5p/pmirGLO-p300 3′ UTR co-transfected group was decreased significantly (P0.05). The protein expression level of p300 was significantly reduced after mir 26b-5p transfection compared with NC-mimic transfection (P0.05)
and the mRNA and protein expression levels of Nanog were significantly down-regulated after the transfection (P0.05). The mir 26b-5p transfection also reduced the proliferation and colony formation of UC-MSCs (P0.05). Conclusion mir 26b-5p suppresses the protein expression levels of p300 and Nanog
a pluripotency transcription factor in UC-MSCs
and also inhibits the cell proliferation and colony formation capability.