Objective: To investigate the influence of GPT2（glutamic pyruvate transaminase 2）to biological characteristics of human acute myeloid leukemia cell line HL-60.
Methods: The expression of GPT2 in hematological tumor and AML cell was detected. The lentvirus-mediated of short-hairpin RNA (shRNA) was constricted, and the knock-down efficiency of HL-60 in AML cell after infected by lentvirus-mediated was detected by Western blot and Q-PCR. CCK-8 assay and soft agar colony formation assay were used to detect the effect of GPT2 gene deletion to the cell proliferation potential. Fluorescence activated cell sorting(FACS) was used to analyze the effect of gene deletion to the cell cycle and Caspase 3/7 Activity Assay Kit was used to analyze the effect of GPT2 gene deletion to the cell apoptosis.
Results: GPT2 showed mRNA high expression in AML patients. CCK-8, soft agar assay, and Caspase 3/7 Activity Assay Kit results showed that compared with shCtrl group, the cells in shGPT2-1、shGPT2-2、shGPT2-3 group showed the slowing down on proliferation, decreasing on colony ability, and the apoptosis of the cells was increasing significantly. FACS showed that GPT2 gene was related to the cycle of HL-60 cell.
Conclusion: GPT2 appears to involve the proliferation, cycle distribution and apoptosis of AML cell HL-60. The deletion of GPT gene can lead to the inhibitation of cells proliferation and increase apoptosis.
题目: 谷丙转氨酶GPT2对急性髓系白血病细胞HL-60生长特性的影响.
目的: 探讨参与三羧酸（TCA）循环中谷氨酰胺代谢的谷丙转氨酶2（glutamic pyruvate transaminase 2,GPT2）对急性髓系白血病细胞生长、增殖及凋亡的影响.
方法: 数据库挖掘了解不同血液肿瘤以急性白血病细胞中GPT2的表达水平，通过构建目的基因short-hairpin RNA (shRNA)慢病毒，转染急性髓系白血病细胞HL-60后验证目的基因敲减效率。采用CCK-8法检测GPT2基因敲减对细胞增殖的影响; 通过软琼脂集落形成实验检测基因敲减对细胞集落形成能力的影响；FACS（fluorescence activated cell sorting）检测基因敲减对细胞周期的影响；Caspase 3/7检测GPT2基因敲减对凋亡的影响.
结果: GPT2在AML患者中存在mRNA水平高表达。CCK-8检测、软琼脂克隆形成实验以及Caspase 3/7检测结果表明，与shCtrl组比较，shGPT2-1、shGPT2-2、shGPT2-3组细胞增殖减缓，集落形成能力减弱，凋亡细胞数显著增加。细胞周期实验结果表明，GPT2基因与HL-60细胞的周期分布显著相关.
结论: GPT2在人AML细胞HL-60中参与肿瘤细胞的增殖、周期分布以及细胞的凋亡。GPT基因的敲减会导致肿瘤细胞的增殖抑制以及凋亡增加.