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目的探讨1,25-二羟维生素D_3[1,25(OH)_2D_3]对人白血病6T-CEM细胞株凋亡及维生素D受体(VDR)蛋白表达的影响。方法培养6T-CEM细胞株,采用随机数字表法将其分为A、B、C、D四组,A、B、C组分别加入浓度为10-6、10-7、10-8mol/L的1,25(OH)_2D_3,D组加入DMSO。采用MTT法检测各组吸光度A值和细胞增殖抑制率情况,AO-EB荧光染色观察各组细胞形态,流式细胞仪检测各组细胞凋亡情况,RT-PCR和Western blot法检测VDR mRNA和蛋白表达。结果 A组吸光度A值明显低于B、C组和D组(P<0.05);A、B、C三组的细胞增殖抑制率从高到低依次排序为A>B>C;D组CEM细胞凋亡指数明显低于A、B、C组(P<0.05);A组凋亡指数明显高于其他三组(P<0.05);各组VDR mRNA表达差异无统计学意义(P>0.05);A组VDR蛋白表达明显高于其他三组(P<0.05)。结论 1,25(OH)_2D_3能有效抑制6T-CEM细胞增殖,诱导其凋亡,上调VDR蛋白表达。
Objective To investigate the effects of 1,25-dihydroxyvitamin D_3 [1,25 (OH) _2D_3] on the apoptosis and the expression of vitamin D receptor (VDR) in human leukemia 6T-CEM cells. Methods Cell lines of 6T-CEM were cultured and divided into A, B, C and D groups by random number table. Groups A, B and C were added with 10-6, 10-7 and 10-8 mol / L Of 1,25 (OH) _2D_3, D group added DMSO. The absorbance A value and inhibition rate of cell proliferation were detected by MTT assay. Cell morphology was observed by AO-EB staining. Cell apoptosis was detected by flow cytometry. VDR mRNA and protein were detected by RT-PCR and Western blot Protein. Results The absorbance A value of group A was significantly lower than that of group B, C and D (P <0.05). The inhibitory rates of cell proliferation in groups A, B and C were A> B> C, The apoptosis index in group A was significantly lower than that in groups A, B and C (P <0.05), and the apoptosis index in group A was significantly higher than that in the other three groups (P <0.05). There was no significant difference in VDR mRNA expression between groups ). The expression of VDR protein in group A was significantly higher than the other three groups (P <0.05). Conclusion 1,25 (OH) _2D_3 can effectively inhibit the proliferation of 6T-CEM cells, induce apoptosis and up-regulate VDR protein expression.