磷脂酶C通过抑制T-ALL细胞凋亡发挥促癌作用  

Potential Oncogenic Role of Genes in PLC1 Family on T-ALL Cell Lines by Suppression of Apoptosis

作　　者：曾腊梅 舒逸[1] 朱丹 马德禹 Filippus I Tshavuka 张虹洋 邹琳[1,2] ZENG La-Mei;SHU Yi;ZHU Dan;MA De-Yu;Filippus I Tshavuka;ZHANG Hong-Yang;ZOU Lin(Department of Clinical Molecular Medicine of Childrens Hospital in Chongqing Medical Universiy,National Clinical Research Center for Child Health and Disorders,Ministry of Education Key Laboratory of Child Development and Disorders,Chongqing Key Laboratory of Peditrics,Chongqing 400014,China;Clinical Research Unit of Children's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Institute of Pediatric Infection,Immunity and Critical Care Medicine,Shanghai Jiao Tong Universiny School of Medicine,Shanghai 200062,China)

机构地区：[1]重庆医科大学附属儿童医院临床分子医学中心,国家儿童健康与疾病临床医学研究中心,儿童发育疾病研究教育部重点实验室,儿科学重庆市重点实验室,重庆400014 [2]上海交通大学医学院附属儿童医院临床研究中心,上海交通大学医学院儿童感染免疫与重症医学研究院,上海200062

出　　处：《中国实验血液学杂志》2023年第4期931-938,共8页Journal of Experimental Hematology

基　　金：国家自然科学基金(82270160,82070167,81900190)。

摘　　要：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,AnnexinV-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、PLCη1siRNA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Westernblot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。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 PLCBI,PLCyI,PLCηI 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 Westem 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 TA RGET-ALL database,the prognosis of T-ALL patients with high expression of PLQ8I,PLCyI and PLCηI was poor,and PLQ3I,PLCyI,PLCηI were unevenly distributed in T-ALL subtypes.PLCB1,PLCyI and PLCηl maintained the survival of P12-ICH and CCRF-CEM cell lines,respectively,while they had no effect on the survival of MOLT4.Conclusion:PLQBI,PLCyI and PLCηl can maintain the growth of T-ALL cell lines in vitro and promote the malignant progression of T-ALL,which are potential therapeutic targets.

关 键 词：磷脂酶C 急性T淋巴细胞白血病 细胞凋亡 预后 

分 类 号：R733.71[医药卫生—肿瘤]