Effects of SHOX2 over-expression on BMP9-induced osteogenic differentiation of mouse mesenchymal stem cell line C3H10T1/2

Abstract

Abstract: Objective To investigate the effects of short stature homeobox 2(SHOX2) on bone morphogenetic protein 9(BMP9)-induced osteogenic differentiation of mouse mesenchymal stem cell line C3H10T1/2. Methods After construction of SHOX2 recombinant adenoviruse (Ad-SHOX2), normal control(NC) group, Ad-SHOX2 group, BMP9 group, as well as BMP9+Ad-SHOX2 group were set up. Early osteogenic ability was evaluated with alkaline phosphatase(ALP); Late osteogenic ability was detected by Ca²⁺ deposition; Cell proliferation was observed using microscope; Meanwhile, Western blot was employed to confirm protein expression of proliferating cell nuclear antigen (PCNA), runt-related transcription factor 2(RUNX2), T-ERK1/2 and p-ERK1/2. Results Ad-SHOX2 was constructed successfully. Ad-SHOX2 and BMP9 groups produced higher ALP activity, more Ca²⁺ accumula-tion, and higher RUNX2 expression in comparison with NC group (P＜0.05). Compared with Ad-SHOX2 and BMP9 groups, ALP activity in BMP9+Ad-SHOX2 group was enhanced (P＜0.05). Ad-SHOX2(low infection)+BMP9 group dampened Ca²⁺ accumulation remarkably. And that with high infection stimulated Ca²⁺ accumulation compared with BMP9 group, but changeless in contrast with Ad-SHOX2 group. Meanwhile, RUNX2 expression in BMP9+Ad-SHOX2 group was higher compared with BMP9 group's, but no change was found in Ad-SHOX2 group (P＜0.05). Cell growth density in Ad-SHOX2 and BMP9 groups was greater than NC group's, consistent with protein level of PCNA(P＜0.05). And protein expression of PCNA in BMP9+Ad-SHOX2 group was increased compared with BMP9 group's(P＜0.05). Additionally, protein expressions of T-ERK1/2 and p-ERK1/2 in both Ad-SHOX2 group and BMP9+Ad-SHOX2 group were up-regulated compared with NC group (P＜0.05). Conclusions SHOX2 may promote proliferation and osteogenic differentiation of C3H10T1/2 cells via regulating MAPK/ERK signaling pathway in the absence of exogenous osteoinductive factors. However, its effects on osteogenic differentiation with BMP9 potentially function with antagonistic effect and this is target for further research.

Key words: short stature homeobox2(SHOX2), bone morphogenetic protein 9(BMP9), osteogenic differentiation, C3H10T1/2 cell

CLC Number:

Q254

ZHANG Ping, YUAN Xiao-hui, HUANG Hua-kun, YANG Chun-mei, ZHANG Lu-lu, LUO Xiao-ji, LUO Jin-yong. Effects of SHOX2 over-expression on BMP9-induced osteogenic differentiation of mouse mesenchymal stem cell line C3H10T1/2[J]. Basic & Clinical Medicine, 2021, 41(1): 1-7.

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