Effects of Sox-9 gene silenced by small interfering RNA on proliferation and apoptosis of epiphysis stem cells

doi:10.3969/j.issn.2095-4344.2013.40.008

Abstract

Abstract:

BACKGROUND: Current research has shown that many cell growth factors can promote the proliferation and differentiation of epiphysis stem cells, and this regulation is closely related to Sox-9.
OBJECTIVE: To investigate the effects of small interfering RNA-induced specific silence of Sox-9 gene on the proliferation and apoptosis of epiphysis stem cells.
METHODS: Epiphysis stem cells were isolated from the ring of La Croix with enzyme digestion and purified with magnetic activated cell sorting, identified by immunocytochemistry assay. The cells in good conditions were selected to be transfected by Sox-9 silenced by small interfering RNA with fluorescent marker, and then were observed under the fluorescent microscope to check the transfection efficiency. Next step, epiphysis stem cells were divided into 2 groups: group one was cultured normally as control group; group two was transfected by Sox-9 silenced by small interfering RNA through Lipofectamine^TM 2000 as experimental group.
RESULTS AND CONCLUSION: The primary epiphysis stem cells were separated and purified, which were of stable growth and tightly attachment. The results of immunohistochemistry and immunofluorescence showed the epiphysis stem cells expressed cell-specific markers, fibroblast growth factor receptor 3. After transfection, reverse transcription-PCR results showed that Sox-9 gene expression in epiphysis stem cell was inhibited specifically; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide results showed that cell proliferation in experimental group decreased significantly compared with the control group (P < 0.05), and the cell apoptosis detected by flow cytometry showed that the experimental group increased as compared with the control group (P < 0.05). Sox-9 gene plays an important role in regulating the proliferation and apoptosis of rat epiphysis stem cells.

Key words: epiphyses, stem cells, biology, chondrocytes, cartilage

CLC Number:

R394.2

Chen Shao-jian, Guo Feng-jin, Wang Chun, Gong Chen. Effects of Sox-9 gene silenced by small interfering RNA on proliferation and apoptosis of epiphysis stem cells　[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(40): 7068-7075.

Figures/Tables 6

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