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| Effect of bone marrow mesenchymal stem cells on the senescence of K562 cells and the Raf/MEK/ERK signaling pathway |
| ZHOU Wen ZHOU Yue TANG Ziyun HE Siyue LIU Xiaohu BI Ziying LI Minrui YANG Nan |
| Teaching and Research Office of Histology and Embryology, Dali University, Yunnan Province, Dali 671000, China |
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Abstract Objective To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) on the senescence of chronic myeloid leukemia K562 cells and Raf/MEK/ERK signaling pathway. Methods BMSCs were isolated from the bone marrow of 30 C57BL/6J male mice with SPF grade of 6-8 weeks and body weight of 20-30 g. The expression of surface antigens CD29, CD244, and CD45 was detected by flow cytometry. After induction with adipogenic and osteogenic differentiation inducing fluid, the differentiation ability was identified by oil red O and alizarin red staining. BMSCs and K562 cells were cultured at the ratios of 1∶4, 1∶2, 1∶1, and 2∶1 for 24, 48, and 72 h, respectively. The co-culture time and cell ratio with the lowest proliferation rate of K562 cells were selected as the BMSCs group, and the conventional cultured K562 cells were used as the control group for subsequent experiments. SA-β-Gal staining was used to detect the senescence of cells in the two groups, and Western blot was used to detect the protein expressions of p16INK4a, p-Raf, p-MEK, and p-ERK in the two groups. Results The positive expression rates of BMSCs surface markers CD29, CD44, and CD45 were 89.18%, 87.65%, and 1.16%, respectively. BMSCs successfully differentiated into adipocytes and osteocytes in vitro. When the ratio of BMSCs to K562 cells was 1∶2, 1∶1, and 2∶1, the proliferation rates of K562 cells at each time point of co-culture were lower than those of the control group at the same time point, and the proliferation rates of K562 cells at each time point with the ratio of 2∶1 were lower than those of the control group at the same time point (P ＜ 0.05). When the ratio of BMSCs to K562 cells was 2∶1, the proliferation rate of K562 cells after co-culture for 48 h was lower than those after co-culture for 24 and 72 h (P ＜ 0.05). The positive rate of senescent cells and the protein expression of p16INK4a in BMSCs group were higher than those in control group, while the protein expression levels of p-Raf, P-MEK, and p-ERK were lower than those in control group (P ＜ 0.01). Conclusion BMSCs can induce senescence in K562 cells, and the mechanism may be related to Raf/MEK/ERK signaling pathway.
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