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目的探讨小鼠增殖相关蛋白p38-2G4基因表达上调对小鼠红白血病(MEL)细胞增殖和诱导分化能力的影响。方法构建p38-2G4-pLJM1高表达重组载体与pCMV-VSV-G和pCMV-dR8.2包装质粒共转染HEK293T细胞慢病毒,感染MEL细胞,建立稳定高表达p38-2G4蛋白的MEL细胞系,Western blotting检测稳定株p38-2G4蛋白的表达。MTT实验和联苯胺染色法检测p38-2G4蛋白高表达对MEL细胞增殖和诱导分化能力的影响。结果高表达稳定株中p38-2G4蛋白表达量明显高于对照组细胞(P<0.05)。p38-2G4蛋白的高表达能显著影响MEL诱导分化过程,导致血红蛋白合成减少(P<0.05),但不能明显改变细胞活力(P>0.05)。结论 p38-2G4蛋白的高表达对MEL细胞增殖无明显影响,但能显著影响其被丁酸钠诱导分化的过程。
Objective To investigate the effects of up-regulation of p38-2G4 gene on proliferation, differentiation and differentiation of murine erythroleukemia (MEL) cells. Methods Recombinant plasmid p38-2G4-pLJM1 was constructed and co-transfected with pCMV-VSV-G and pCMV-dR8.2 plasmids into HEK 293T lentivirus to infect MEL cells. The MEL cell line stably expressing p38-2G4 protein was established. The expression of p38-2G4 protein was detected by Western blotting. MTT assay and benzidine staining were used to detect the effect of p38-2G4 overexpression on the proliferation and differentiation of MEL cells. Results The expression of p38-2G4 protein in the highly expressed and stable strains was significantly higher than that in the control group (P <0.05). High expression of p38-2G4 protein significantly affected the process of MEL induced differentiation, resulting in decreased hemoglobin synthesis (P <0.05), but not significantly change cell viability (P> 0.05). Conclusion The high expression of p38-2G4 protein has no significant effect on the proliferation of MEL cells, but it can significantly affect the induction of differentiation by sodium butyrate.