Study On Expression Of ERK Signal Pathway In Bone Marrow Nucleated Red Blood Cells Of Patients With CMS

Purpose By observing the CMS nucleated red blood cells in the bone marrow of patients Ras, Raf, MEK, ERK level change, and to explore ERK signaling pathway in CMS occurrence and development role. Methods Select male CMS patients and simple old fracture patients all 15 cases from Affiliated Hospital of Qinghai University, collected bone marrow fluid, isolated bone marrow mononuclear cells with CD71 and CD235 a antibody screening positive cells. Bone marrow nucleated red blood cells of CMS were divided into five groups: control group, DMSO solvent control group, PD98059 5?mol/L, PD98059 10?mol/L, PD98059 20?mol/L.The cells were cultured 72 h in 37°C, 5% CO2, 3% O2 incubator, by Elisa method to measure the Ras-GTP levels in supernatant, using RT-PCR method to determine the expression of BRaf, MEK, ERK1, ERK2 mRNA in nucleated red blood cells. and Western Blot method to detect expression of BRaf, MEK, ERK1/2 protein. Results 1 Ras-GTP levels in marrow nucleated red blood cells of CMS group [(21.34?0.78) pg/mL] was higher than the control group [(19.38?0.88) pg/mL], the difference was statistically significant(P <0.05). CMS group BRaf, MEK, ERK1/2 mRNA levels(1.60?0.38, 2.57?1.50, 1.11?0.59, 1.08?0.79) were higher than the control group(0.81?0.44, 0.95?0.41, 0.52?0.55, 0.38?0.28), the difference was statistically significant(P <0.05). 2 Effect of PD98059 on Ras-GTP: After PD98059 treated with different concentrations of the bone marrow nucleated red blood cells, ELISA detect level of Ras-GTP, found no significant difference among five groups of Ras-GTP(P = 0.798). 3 Effect of PD98059 on BRaf, MEK and ERK1/2 mRNA: Compared with the control group, the expression of BRaf, MEK mRNA was not statistically significant(P values were 0.826, 0.298). Compared with the PD98059 20?mol / L group, the expression level of ERK1/2 mRNA was statistically significant(P = 0.001, 0.002).4 Effect of PD98059 on p-BRaf, p-MEK, p-ERK1/2 protein: Compared with the control group, expression of p-BRaf, p-MEK protein was not statistically significant(P values were 0.370, 0.351). Compared with the PD98059 20?mol / L group, the difference of p-ERK1/2 protein level was statistically significant(P values were <0.001, 0.007). Conclusions 1 Ras/Raf/MEK/ERK pathway may be involved in abnormal activation of the CMS excessive accumulation of red blood cells, suggesting that it may be one mechanism of signal transduction pathways in the happen of CMS. 2 PD98059 can cut the expression of ERK1 / 2 mRNA and p-ERK1 / 2 proteins.