Objective: To explore the role of phospholipase C(PLC) family in the progression of acute T lymphoblastic leukemia (T-ALL).
Methods: The apoptosis of T-ALL cells was determined by Annexin V-PE/7-AAD staining after treatment of PLC inhibitor U73122 and Edelfosine. Cox regression and Kaplan-Meier were used to analyze the impact of PLC expressions on the event-free survival (EFS) of T-ALL patients. PLC expression in each subtype of T-ALL were analyzed by One-way ANOVA. The siRNA expression plasmids targeting the PLCβ1, PLCγ1, PLCη1 gene were constructed, and T-ALL cells were infected with retrovirus packaging in HEK-293T cells. The mRNA and protein level were tested by RT-PCR and Western blot.
Results: P12-ICH and CCRF-CEM cell line were sensitive to U73122 and Edelfosine treatment, while Jurkat and MOLT4 were resistant to them. In the TARGET-ALL database, the prognosis of T-ALL patients with high expression of PLCβ1, PLCγ1 and PLCη1 was poor, and PLCβ1, PLCγ1, PLCη1 were unevenly distributed in T-ALL subtypes. PLCβ1, PLCγ1 and PLCη1 maintained the survival of P12-ICH and CCRF-CEM cell lines, respectively, while they had no effect on the survival of MOLT4.
Conclusion: PLCβ1, PLCγ1 and PLCη1 can maintain the growth of T-ALL cell lines in vitro and promote the malignant progression of T-ALL, which are potential therapeutic targets.
题目: 磷脂酶C通过抑制T-ALL细胞凋亡发挥促癌作用.
目的: 探讨磷脂酶C(PLC)家族各成员在急性T淋巴细胞白血病（T-ALL）进展中的作用.
方法: 泛PLC抑制剂U73122和Edelfosine处理T-ALL细胞，Annexin V-PE/7-AAD双染法检测细胞凋亡。Cox回归和Kaplan-Meier法分析PLC蛋白对T-ALL患者无事件生存率（EFS）的影响。单因素方差分析PLC在各亚型T-ALL中的表达。构建PLCβ1、PLCγ1、 PLCη1 siRNA逆转录质粒，HEK-293T细胞包装逆转录病毒感染T-ALL细胞。利用RT-PCR和Western blot检测基因表达.
结果: 4种T-ALL细胞系中，P12-ICH和CCRF-CEM对 U73122和Edelfosine敏感，Jurkat和MOLT4对其耐药。TARGET-ALL数据库中，PLCβ1、PLCγ1、 PLCη1高表达T-ALL患者预后不良，PLCβ1、PLCγ1、 PLCη1在T-ALL各亚型中分布不均。在P12-ICH和CCRF-CEM中，PLCβ1、PLCγ1和 PLCη1分别维持白血病细胞生存，而在MOLT4中，PLCβ1、PLCγ1和 PLCη1对细胞生存无影响.
结论: PLCβ1、PLCγ1和 PLCη1能维持T-ALL细胞株体外生长，促进T-ALL恶性进展，是潜在的治疗靶点.
Keywords: acute T lymphoblastic leukemia; apoptosis; phospholipase C; prognosis.