Expression of microRNA in human bone marrow-derived mesenchymal stem cells
- VernacularTitle:miRNAs在人骨髓间充质干细胞中的表达
- Author:
Jie GAO
;
Tongtao YANG
;
Xiuchun QIU
- Publication Type:Journal Article
- Keywords:
bone marrow;
mesenchymal stem cells;
cells , cultured;
microRNAs
- From:
Medical Journal of Chinese People's Liberation Army
1982;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To identify the expression of miRNAs in human bone marrow-derived mesenchymal stem cells (hMSCs). Method Low molecular weight RNA fraction (≤200nt) from human bone marrow-derived mesenchymal stem cells was extracted, and polyadenylated by poly(A) polymerase. Then a 5′RNA adapter was ligated to poly(A)-tailed RNA using T4 RNA ligase. RNAs were reversely transcribed and amplified by RT-PCR. The PCR products with a size of about 109 bp were recovered and cloned into pCR 4-TOPO vector. After sequencing, database searching, and expression profiling, the expression of miRNAs in hMSCs was sieved. The expression of novel and some known miRNAs was examined by Northern blotting with small RNAs (≤200nt) isolated from hMSCs, osteosarcoma cell line SOSP-9607 and UMR-106. Results hMSCs were isolated from bone marrow and purified by centrifuge and in vitro culture. Cell markers CD44 were positive, but CD34 and CD45 were negative. A total of 194 clones were subsequently characterized by DNA sequencing and database searching. 52 clones (correspond to 27species) out of 194 clones from hMSCs were identified as miRNAs. One novel miRNAs (PREDICTED-miR-202) was discovered among other 26 known miRNAs, which were predicted in Nature. Northern blotting confirmed that 1 novel miRNAs and 3 known miRNAs (miR-495, miR-34a, miR-17-5p, PREDICTED-miR-202) were stably expressed in hMSCs. Conclusion These findings indicate that a large diverse population of miRNAs are likely important regulators for hMSCs in maintaining their state of self-renewal, and have potential value in stem cells research.
