Lentivirus-mediated over-expression of beta-catenin accelerates proliferation and migration of mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2013.40.009

Abstract

Abstract:

BACKGROUND: β-catenin is the most critical signaling molecule in the Wnt/β-catenin signaling pathway, which is involved in the regulation of cell proliferation, differentiation and tissue self-healing balance.
OBJECTIVE: To construct a stable β-catenin over-expression lentivirus-mediated vector and to transfect mesenchymal stem cells line for investigating its effects on proliferation and migration of mesenchymal stem cells.
METHODS: Over-expression vector, PLV-EF1A-catenin-RFP, was constructed and transfected the 293T cell to infect mesenchymal stem cells, and positive cells were selected with puromycin. The up-regulated efficiency of targeting β-catenin gene at mRNA level was detected by real-time quantitative PCR, the effect on proliferation of mesenchmal stem cell was assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve, and the migration ability was detected by Transwell motility assay.
RESULTS AND CONCLUSION: The lentiviral vector targeting β-catenin gene was constructed successfully, and a stable mesenchymal stem cell line that up-regulated β-catenin was established. Real-time quantitative PCR results showed that the expression of β-catenin gene was efficiently up-regulated by infecting PLV-EF1A-catenin-RFP (P < 0.05). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve showed that cell doubling time was shortened after infected with pLV-EF1A-catenin-RFP (P < 0.05), indicating that the over-expression of the β-catenin gene successfully increased the proliferative capability of mesenchymal stem cells. The Transwell assay also showed similar increasing results on the migration ability (P < 0.01). The lenvivirus-mediated over-expression of the β-catenin gene can be used to increase the proliferation and migration abilities of the mesenchymal stem cells.

Key words: mesenchymal stem cells, lentivirus infections, beta Catenin, cell proliferation, cell movement

CLC Number:

R394.2

Wu Qian, Wei Ya-ming, Li Yu-yuan, Nie Yu-qiang, Cao Yan-wen. Lentivirus-mediated over-expression of beta-catenin accelerates proliferation and migration of mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(40): 7076-7083.

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