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Impacts of K562 cells towards activities of Toll-like receptor pathway of human mesenchymal stem cell-bone marrow.

ABSTRACT: The study aim was to investigate the impacts of K562 cells towards the activities of Toll-like receptor pathway of human mesenchymal stem cell-bone marrow (HMSC-bm). The in vitro co-culture of HMSC-bm and K562 cells was set as the experiment group (HMSC-bm + K562), the HMSC-bm cultured alone was set as the control group (HMSC-bm), the expressions of six interested genes and their proteins, namely MyD88, P38, NF-?B, TAB1, TLR3 and TBK1, of the Toll -like receptor signaling pathway were detected and compared, as well as the secretions of such cytokines as IL-6, IL-8, TNF-? and IFN-? in the cell supernatant, which were regulated by the Toll-like receptor pathway. The expressions of MyD88, P38, TAB1 and TLR3 of the HMSC-bm + K562 group were higher than the HMSC-bm group, while that of TBK1 was lower, and the NF-?B expression showed no significant difference between the two groups (P > 0.05). Compared with the HMSC-bm group, the supernatant of HMSC-bm + K562 group exhibited the higher secretion levels of IL-6 and IL-8, while that of IFN-? was just contrary, and the differences were significant (P < 0.05). The secretion levels of TNF-? within the two groups were not significantly different (P > 0.05). The co-culture of K562 and HMSC-bm could induce the activity changes of Toll-like receptor pathway of HMSC-bm, which was beneficial towards the proliferation of K562 cells.

SUBMITTER: Xu Z

PROVIDER: S-EPMC4537973 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Impacts of K562 cells towards activities of Toll-like receptor pathway of human mesenchymal stem cell-bone marrow.

Xu Zheng Z Luo Jianmin J Yang Lin L Wang Xingzhe X Pan Yuxia Y Shang Yintao Y Yang Jingci J

International journal of clinical and experimental medicine 20150615 6

The study aim was to investigate the impacts of K562 cells towards the activities of Toll-like receptor pathway of human mesenchymal stem cell-bone marrow (HMSC-bm). The in vitro co-culture of HMSC-bm and K562 cells was set as the experiment group (HMSC-bm + K562), the HMSC-bm cultured alone was set as the control group (HMSC-bm), the expressions of six interested genes and their proteins, namely MyD88, P38, NF-κB, TAB1, TLR3 and TBK1, of the Toll -like receptor signaling pathway were detected a ...[more]

PMID: 26309591

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Project description:Chordin-like 1 (CHRDL1) is a secreted glycoprotein with repeated cysteine-rich domains, which can bind to BMPs family ligands. Although it has been reported to play important roles in several systems, the exact roles of CHRDL1 on human bone mesenchymal stem cells (hBMSCs) osteogenesis remain to be explored. The present study aimed to investigate the roles of CHRDL1 on the osteogenic differentiation of hBMSCs and the underlying molecular mechanisms. We found that CHRDL1 was upregulated during hBMSCs osteogenesis, and rhBMP-4 administration could enhance CHRDL1 mRNA expression in a dose and time dependent manner. Knockdown of CHRDL1 did not affect hBMSCs proliferation, but inhibited the BMP-4-dependent osteogenic differentiation, showing decreased mRNA expression levels of osteogenic markers and reduced mineralization. On the contrary, overexpression of CHRDL1 enhanced BMP-4 induced osteogenic differentiation of hBMSCs. Moreover, in vivo experiments by transplanting CHRDL1 gene modified hBMSCs into nude mice defective femur models displayed higher new bone formation in CHRDL1 overexpression groups, but lower new bone formation in CHRDL1 knockdown groups, compared with control groups. In consistent with the bone formation rate, there were increased CHRDL1 protein expression in new bone formation regions of defective femur in CHRDL1 overexpression groups, while reduced CHRDL1 protein expression in CHRDL1 knockdown groups compared with control groups. These indicate that CHRDL1 can promote osteoblast differentiation in vivo. Furthermore, the mechanisms study showed that CHRDL1 improved BMP-4 induced phosphorylation of SMAD1/5/9 during osteogenic differentiation of hBMSCs. Besides, promotion of osteogenic differentiation and the activation of SMAD phosphorylation by CHRDL1 can be blocked by BMP receptor type I inhibitor LDN-193189. In conclusion, our results suggested that CHRDL1 can promote hBMSCs osteogenic differentiation through enhancing the activation of BMP-4-SMAD1/5/9 pathway.

Dataset Information

Impacts of K562 cells towards activities of Toll-like receptor pathway of human mesenchymal stem cell-bone marrow.

Publications

Impacts of K562 cells towards activities of Toll-like receptor pathway of human mesenchymal stem cell-bone marrow.

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