Jichu yixue yu linchuang (Jan 2021)

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

  • ZHANG Ping, YUAN Xiao-hui, HUANG Hua-kun, YANG Chun-mei, ZHANG Lu-lu, LUO Xiao-ji, LUO Jin-yong

Journal volume & issue
Vol. 41, no. 1
pp. 1 – 7

Abstract

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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.

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