ARHI基因抑制U937白血病细胞株生长并诱导其G_2/M期阻滞及凋亡

目的:探讨抑癌基因ARHI在急性髓细胞白血病中的表达情况,同时探索其对U937细胞生长的影响。方法:RT-PCR方法检测急性髓细胞白血病细胞株、人胚肾细胞293FT、白血病原代细胞以及健康志愿者血细胞中ARHI mRNA的表达情况。构建高表达ARHI的MSCV-IRES-GFP-ARHI质粒转染U937细胞,MTT法连续8 d检测细胞活性绘制生长曲线。新鲜培养基重悬U937细胞24 h后,采用流式细胞术检测细胞周期以及细胞凋亡。结果:293FT细胞、健康志愿者血细胞中均检测到ARHI mRNA表达,而白血病细胞株以及白血病患者原代细胞中未检测到该基因的表达。生长曲线显示高表达ARHI基因的U937(U937-ARHI)细胞在第6~8天细胞活力低于对照(U937-GFP)组。高表达ARHI的U937细胞G2/M期细胞比例及细胞凋亡率均高于对照组(P0.05)。结论:ARHI在急性髓系白血病细胞中的表达减低;高表达ARHI基因能抑制U937细胞生长、阻滞其细胞周期在G2/M期并促进细胞凋亡。

ARHI gene inhibits cell growth,induces G2/M phase arrest and apoptosis of acute myeloid leukemia cell line U937

AIM: To investigate the expression of aplasia rashomolog member I (ARHI) gene in acute myeloid leukemia cells (AML) and to study the effects of ARHI on the growth of AML cell line U937.METHODS: The mRNA expression of ARHI in AML cells, 293FT cells, AML primary cells and healthy volunteer blood cells were detected by RT-PCR. After transfection with the MSCV-IRES-GFP-ARHI plasmid to the U937 cells, the growth curve was analyzed by MTT assay. U937 cells were re-suspended by fresh medium and cultured for 24 h, then the cell cycle distribution and apoptotic rate were determined. RESULTS: The mRNA of ARHI was positively detectable in 293FT cells and healthy volunteer blood cells instead of AML cell line and AML primary cells. The growth curve showed that cell viability in U937 cells with high expression of ARHI (U937-ARHI) was lower than that in the control cells (U937-GFP) on 6th~8th day. The ratio of G₂/M phase and apoptotic rate in the U937-ARHI cells were increased compare with control group(P<0.05). CONCLUSION: The mRNA level of ARHI is low in AML cells. High expression of ARHI gene in U937 cells inhibits cell growth, arrests the cells at G₂/M phase and induces apoptosis.