过表达转录因子BACH2对人急性淋巴细胞白血病T淋巴细胞活力及凋亡的影响

Effect of transcription factor BACH2 over-expression on viability and apoptosis of human acute lymphoblastic leukemia T lymphocytes

目的:探讨BTB与CNC同源基因2(BACH2)过表达对人急性淋巴细胞白血病T淋巴细胞CCRFCEM活力和凋亡的作用。方法:将人急性淋巴细胞白血病T淋巴细胞CCRF-CEM分为3组:对照组、空载体组和BACH2过表达组,通过脂质体转染法将BACH2过表达载体转入BACH2过表达组的CCRF-CEM细胞。采用qPCR检测CCRF-CEM细胞和正常人外周血单个核细胞(PBMC)中BACH2的mRNA表达差异,CCK8法检测CCRF-CEM细胞活力,流式细胞术检测细胞凋亡水平,免疫荧光染色观察细胞中BACH2和细胞周期蛋白D3(cyclin D3)表达的变化,Western blot检测细胞中cyclin D3、Bcl-2及caspase-3的蛋白表达水平。结果:CCRF-CEM细胞中BACH2的mRNA表达水平显著低于PBMC(P<0.05)。与对照组比较,BACH2过表达组CCRF-CEM细胞活力显著减弱(P<0.05),凋亡率显著升高(P<0.05),cyclin D3和Bcl-2表达水平显著降低(P<0.05),而caspase-3表达水平显著升高(P<0.05)。结论:人急性淋巴细胞白血病T淋巴细胞中BACH2呈低表达,过表达BACH2可抑制人急性淋巴细胞白血病T淋巴细胞的活力,并诱导其凋亡。

AIM:To investigate the effect of over-expression of BTB and CNC homology 2(BACH2)on the vi⁃ability and apoptosis of human acute lymphoblastic leukemia T lymphocytes CCRF-CEM.METHODS:CCRF-CEM cells were divided into 3 groups:control group,empty vector group,and BACH2 over-expression group.The BACH2 over-ex⁃pression vector was transfected into CCRF-CEM cells of BACH2 over-expression group by liposome transfection method.The difference in mRNA expression of BACH2 between CCRF-CEM cells and peripheral blood mononuclear cell(PBMC)was detected by qPCR.CCK8 assay was performed to evaluate the viability of CCRF-CEM cells.Flow cytometry was used to analyzed the apoptosis of CCRF-CEM cells.The protein expression of BACH2 and cyclin D3 in the CCRF-CEM cells was observed by immunofluorescence staining.The protein expression of cyclin D3,Bcl-2 and caspase-3 was determined by Western blot.RESULTS:The mRNA expression of BACH2 in CCRF-CEM cells was significantly lower than that in PBMC(P<0.05).Compared with control group,BACH2 over-expression significantly suppressed the viability,increased the apoptotic rate and caspase-3 expression,and decreased the expression of cyclin D3 and Bcl-2 in CCRF-CEM cells(P<0.05).CONCLUSION:BACH2 expression is decreased in T lymphocytes of human acute lymphoblastic leukemia.Over-expression of BACH2 inhibited the viability of human acute lymphoblastic leukemia T lymphocyte and induced apop⁃tosis.