Department of Hematology, Shanghai East Hospital, School of Medicine, Tongji University.
Department of Oncology, Naval Medical Center of Chinese People's Liberation Army.
Anticancer Drugs. 2024 Nov 1;35(10):902-911. doi: 10.1097/CAD.0000000000001642. Epub 2024 Jul 22.
Multiple myeloma, which is a clonal plasma cell tumor, derives from a postmitotic lymphoid B-cell lineage and remains untreatable. Group XVI phospholipase A2 (PLA2G16) can either be a tumor suppressor or an oncogene in different types of cancer. This study was intended to explore the role of PLA2G16 in multiple myeloma and to reveal the reaction mechanism. The mRNA and protein expressions of PLA2G16 in human bone marrow stromal cell line HS-5 and multiple myeloma cells were assessed using reverse transcription-quantitative PCR and western blot. The transfection efficacy of sh-PLA2G16 and oe-YAP was examined using reverse transcription-quantitative PCR and western blot. Through cell counting kit-8 assay and 5-ethynyl-2'- deoxyuridine staining, multiple myeloma cell viability and proliferation were detected. Flow cytometry was used to measure cell apoptosis and cell cycle distribution. Oxygen consumption rate, the activities of mitochondrial respiratory chain complexes I-V, and the activity of caspase-3 were estimated with Seahorse XF24 analyzer, oxidative phosphorylation activity assay kit, and caspase-3 assay kit, respectively. Lactate production and glucose consumption were evaluated usingcorresponding assay kits. Western blot was employed to meaure proteins associated with cell cycle, glycolysis, pentose phosphate pathway as well as Hippo/YAP signaling pathway. In this study, PLA2G16 expression was greatly increased in multiple myeloma cells and PLA2G16 silence inhibited cell proliferation, promoted cell apoptosis, facilitated cell cycle arrest, and suppressed the reprogramming of glucose metabolism in multiple myeloma. It was also identified that PLA2G16 depletion inhibited the Hippo/YAP signaling pathway. Further experiments revealed that the overexpression of YAP partially reversed the inhibitory effects of PLA2G16 silence on multiple myeloma cell malignant development and the reprogramming of glucose metabolism. Collectively, PLA2G16 silence impeded multiple myeloma progression and inhibited glucose metabolism reprogramming by blocking the Hippo/YAP signaling pathway.
多发性骨髓瘤是一种克隆性浆细胞肿瘤，来源于有丝分裂后的淋巴 B 细胞系，目前仍然无法治愈。第 XVI 组磷脂酶 A2（PLA2G16）在不同类型的癌症中既可以是肿瘤抑制因子，也可以是癌基因。本研究旨在探讨 PLA2G16 在多发性骨髓瘤中的作用，并揭示其反应机制。采用逆转录定量 PCR 和 Western blot 检测人骨髓基质细胞系 HS-5 和多发性骨髓瘤细胞中 PLA2G16 的 mRNA 和蛋白表达。采用逆转录定量 PCR 和 Western blot 检测 sh-PLA2G16 和 oe-YAP 的转染效果。通过细胞计数试剂盒-8 检测和 5-乙炔基-2'-脱氧尿苷染色检测多发性骨髓瘤细胞活力和增殖。通过流式细胞术检测细胞凋亡和细胞周期分布。采用 Seahorse XF24 分析仪、氧化磷酸化活性测定试剂盒和 caspase-3 测定试剂盒分别测定氧耗率、线粒体呼吸链复合物 I-V 的活性和 caspase-3 的活性。利用相应的测定试剂盒评估乳酸生成和葡萄糖消耗。采用 Western blot 法检测与细胞周期、糖酵解、戊糖磷酸途径以及 Hippo/YAP 信号通路相关的蛋白。本研究中，多发性骨髓瘤细胞中 PLA2G16 的表达显著增加，沉默 PLA2G16 可抑制细胞增殖，促进细胞凋亡，促进细胞周期停滞，并抑制多发性骨髓瘤中葡萄糖代谢的重编程。还发现 PLA2G16 耗竭抑制 Hippo/YAP 信号通路。进一步的实验表明，YAP 的过表达部分逆转了 PLA2G16 沉默对多发性骨髓瘤细胞恶性发展和葡萄糖代谢重编程的抑制作用。总之，PLA2G16 沉默通过阻断 Hippo/YAP 信号通路阻碍多发性骨髓瘤的进展，并抑制葡萄糖代谢重编程。
