Effects of Leukaemia Inhibitory Factor Receptor on the Early Stage of Osteogenic Differentiation of Human Bone Marrow Mesenchymal Cells

Wang, Tao; Yan, R. Q.; Xu, X. Y.; Cao, L. L.; Liu, J. Y.; Zheng, Meirong; Li, Weidong

doi:10.14712/fb2018064050186

Folia Biologica > Archive > 2018, Vol. 64 > Issue 5-6 > Effects of Leukaemia Inhibitory Factor…

Fol. Biol. 2018, 64, 186-194

https://doi.org/10.14712/fb2018064050186

Effects of Leukaemia Inhibitory Factor Receptor on the Early Stage of Osteogenic Differentiation of Human Bone Marrow Mesenchymal Cells

Tao Wang¹, R. Q. Yan², X. Y. Xu¹, L. L. Cao³, J. Y. Liu¹, Meirong Zheng¹, Weidong Li¹

¹Key Laboratory of System Bio-medicine of Jiangxi Province, Jiujiang University, Jiujiang, Jiangxi, People’s Republic of China
²Clinical Skills Center, Affiliated Hospital of Jiujiang University, Jiujiang, Jiangxi, People’s Republic of China
³Department of Endocrinology, Jiujiang Affiliated Hospital of Nanchang University, Jiujiang Jiangxi, People’s Republic of China

Received August 2018
Accepted November 2018

Leukaemia inhibitory factor (LIF) has a wide variety of biological activities. While recent studies have focused on the role of LIF in osteoblast differentiation, the exact role of LIFR during the early stage of osteogenic differentiation remains unclear. We observed that LIFR expression gradually decreased during the early stage of osteogenic differentiation of hMSCs. To evaluate how LIFR regulates osteogenic differentiation in greater depth, we transfected hMSCs with LIFR overexpression and siRNA lentiviral plasmids. Cells were divided into four groups: a negative overexpression control group, a LIFR overexpression group, a negative siRNA control group, and a LIFR siRNA group. On different days (0, 3, and 6) of the osteogenic differentiation of hMSCs, alkaline phosphatase (ALP) activity was assayed with an ALP staining and activity assay kit. Cells were harvested to assess the mRNA and protein expression of LIF, LIFR, and osteogenesis-related factors (ALP; RUNX2; osteonectin) by qRT-PCR and western blot analyses, respectively. In addition, culture supernatants were tested for the LIF content by ELISA. Our results showed that overexpression of LIFR significantly suppressed the osteoblast differentiation of hMSCs. In contrast, LIFR siRNA markedly improved this osteoblast differentiation as determined by ALP staining and activity measurements. Moreover, RUNX2, ALP, and ONN expression was also significantly changed by altering LIFR expression. We further analysed the expression of LIF and LIFR, revealing consistent LIF and LIFR trends during the osteogenic differentiation of hMSCs. Together, these results suggested that LIFR may be a novel negative regulator during the early stage of hMSC osteogenic differentiation.

Keywords

leukaemia inhibitory factor receptor, osteogenic differentiation, human bone marrow mesenchymal stem cells.

Funding

The study was supported by the National Natural Science Foundation of China (Nos. 81460221 and 81560367) and Jiangxi Province Natural Science Foundation of China (No. 20161BAB205197).