Objective: To investigate the effect of β-arrestin1 gene on senescence of T-ALL cells and its possible mechanism.
Methods: The bone marrow specimens of T-ALL patients and controls were collected, the expression of β-arrestin1 and β-arrestin1 in the T-ALL patients was detected by RT-PCR and Western blot, respectively, and the relation of β-arrestin1 expression with the clinical pathologic characteristics and the prognosis of T-ALL patients was analyzed statistically. The stable Jurkat cell line with knocked down or overexpressed β-arrestin1 was constructed, the CCK method was used to detect the Jurkat cell number, the β-gal staining was used to analyze the effect of β-arrestin1 on senescence of Jurkat cells, the cross analysis of RNA-Seg data and KEGG data was performed for screening the possible signaling pathway, and Western blot was performed for varifying the key sites of signaling pathway.
Results: The β-arrestin1 expression in specimens of T-ALL patients decreased (P＜0.01), moreover the β-arrestin1 expression negatively related with peripheral blood cell number (r=-0.601), the blasts in peripheral blood (r=-0.516) and extramedullary infiltration (r=-0.359), while positively related with the response to chemotherapy (r=0.393). The detection of stable Jurkat cell line with knocked-down and overexpressed β-arrestin1 found that the β-arrestin 1 could decrease the Jurkat cell number and accelarate the senescence of Jurkat cells (P＜0.05). The cross analysis of RNA-Seg data and KEGG data showed that the senescence of T-ALL cells may be regulated via RAS-P16-PRb-E2F1 by β-arrestin 1. Western bolt confirmed that β-arrestin1 promoted the expression of Ras and p16, and decreased the expression of pRB and E2F1 (P＜0.05).
Conclusions: β-arrestin1 accelerates the senescence of Jurkat cells via Ras-p16-pRb-E2F1, and delays the progression in T-ALL, which may provide a new hypothesis for the pathogenesis of T-ALL.
题目: β-arrestin 1促进急性T淋巴细胞白血病Jurkat细胞衰老.
目的: 探讨β-arrestin1对急性T淋巴细胞白血病（T-ALL）细胞衰老的作用及其可能机制.
方法: 收集T-ALL患者骨髓样本和对照样本，应用RT-PCR和Western blot检测β-arrestin1在T-ALL患者中的表达，统计分析β-arrestin1的表达与T-ALL患者临床病理特征及预后的关系。构建稳定敲低或过表达β-arrestin1 Jurkat细胞株，应用CCK-8检测Jurkat细胞数，β-半乳糖苷酶染色（SA-β-gal）法检测β-arrestin1对Jurkat细胞衰老的作用，应用RNA-Seq与KEGG数据交叉分析筛选可能的信号通路，Western blot验证信号通路关键位点.
结果: β-arrestin1在T-ALL患者样本中表达降低 （P＜0.01），且β-arrestin1的表达与外周血白细胞数（r=-0.601）、外周血原始细胞数（r=-0.516） 和髓外浸润（r=-0.359）呈负相关，与化疗敏感性（r=0.393）呈正相关。利用敲低或过表达β-arrestin1的稳定Jurkat细胞株发现，β-arrestin1降低Jurkat细胞数（P＜0.05），促进Jurkat细胞衰老（P＜0.05）。RNA-Seq与KEGG数据交叉分析发现，T-ALL患者中β-arrestin1可能通过Ras-p16-pRb-E2F1调控细胞衰老。经Western blot验证发现，β-arrestin1促进Ras和p16的表达，抑制pRB和E2F1的表达 （P＜0.05）.
结论: β-arrestin1可能通过Ras-p16-pRb-E2F1促进T-ALL细胞衰老，延缓T-ALL疾病进程，这为阐释T-ALL新的发病机制提供理论依据.