Kong Qian, Xu Lu-Hong, Xu Wei, Fang Jian-Pei, Xu Hong-Gui
Guangdong Provincial Key Laboratory of Malignant Tumour Epigenetics and Gene Regulation, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, P.R. China.
Department of Pediatrics, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, P.R. China.
Int J Oncol. 2015 Jul;47(1):161-70. doi: 10.3892/ijo.2015.2985. Epub 2015 May 4.
Acute lymphoblastic leukaemia (ALL) is a common paediatric cancer and is among the most curable cancers. However, the acquisition of drug resistance is a significant obstacle to the achievement of favourable outcomes, and autophagy is regarded as a mechanism that underlies chemoresistance. In this study, RT-qPCR was used to measure the expression of HMGB1 and Beclin1 in bone marrow mononuclear cells. A CCK-8 test was conducted to assess cell viability. Western blot, immunofluorescence and transmission electron microscopic analyses were performed to evaluate the autophagy levels. Immunoprecipitation analysis was performed to detect protein-protein interactions in the autophagy complexes. We found that HMGB1 expression correlated with the clinical status of ALL. In vitro, anticancer agent-induced cytotoxic effects were associated with autophagy-related drug resistance, and these effects were ameliorated by FIP200 depletion or the application of autophagy inhibitors. Moreover, the Ulk1‑Atg13-FIP200 complex, which promotes HMGB1 trafficking, acted upstream of the HMGB1-Beclin1 and PI3KC3-Beclin1 complexes and played a critical role in autophagy. Targeting the transformation of autophagic complexes or HMGB1 translocation may suppress autophagy and consequently overcome chemoresistance in leukaemia.
急性淋巴细胞白血病(ALL)是一种常见的儿科癌症,也是最可治愈的癌症之一。然而,耐药性的产生是实现良好预后的重大障碍,自噬被认为是化疗耐药的一种机制。在本研究中,采用RT-qPCR检测骨髓单个核细胞中HMGB1和Beclin1的表达。进行CCK-8试验以评估细胞活力。采用蛋白质免疫印迹法、免疫荧光法和透射电子显微镜分析法评估自噬水平。进行免疫沉淀分析以检测自噬复合物中的蛋白质-蛋白质相互作用。我们发现HMGB1表达与ALL的临床状态相关。在体外,抗癌药物诱导的细胞毒性作用与自噬相关的耐药性有关,FIP200缺失或应用自噬抑制剂可改善这些作用。此外,促进HMGB1转运的Ulk1-Atg13-FIP200复合物在HMGB1-Beclin1和PI3KC3-Beclin1复合物的上游起作用,并在自噬中起关键作用。靶向自噬复合物的转变或HMGB1易位可能抑制自噬,从而克服白血病的化疗耐药性。