To investigate the cellular mechanisms of multiple myeloma (MM), we used liquid chromatography‐tandem mass spectrometry for proteomics analysis of CD138⁺ plasma cells from patients with MM and healthy controls. We found that the 60‐kDa heat shock protein (HSP60, also known as HSPD1) was significantly upregulated in myeloma cells. HSP60 is an important chaperone protein that regulates the homeostasis of mitochondrial proteins and maintains mitochondrial function. Knockdown (KD) of HSP60 in myeloma cells resulted in inhibition of proliferation and reduced the quality of the mitochondria. Mitochondrial stress tests showed that HSP60 KD inhibited glycolysis and mitochondrial activity. Metabolomics showed a decrease in glycolysis and tricarboxylic acid cycle metabolites, and inhibited the formation of creatine and phosphocreatine by the reaction of S‐adenosylmethionine (SAM) with amino acids mediated by demethyladenosine transferase 1, mitochondrial (TFB1M) and reduced energy storage substances. Moreover, HSP60 silencing influenced the synthesis of ribonucleotides and nicotinamide adenine dinucleotide phosphate (NADPH) by the pentose phosphate pathway to inhibit cell proliferation. HSP60 KD inhibited 5' adenosine monophosphate‐activated protein kinase (AMPK), which inhibited the key enzyme 6‐phosphofructo‐2‐kinase/fructose‐2,6‐biphosphatase 3 (PFKFB3), effecting the metabolism of fatty acids by inhibiting malonyl‐coenzyme A. Our data suggest that reduced HSP60 expression alters metabolic reprogramming in MM, inhibits tumour progression and reduces mitochondrial‐dependent biosynthesis, suggesting that HSP60 is a potential therapeutic target for MM treatment.

中文翻译：

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60 kDa热激蛋白调节能量重排和蛋白质合成，从而促进多发性骨髓瘤细胞的增殖。

要研究多发性骨髓瘤（MM）的细胞机制，我们使用液相色谱-串联质谱法对CD138 ⁺进行蛋白质组学分析MM患者和健康对照组的浆细胞。我们发现，骨髓瘤细胞中60 kDa的热激蛋白（HSP60，也称为HSPD1）明显上调。HSP60是一种重要的伴侣蛋白，可调节线粒体蛋白质的体内平衡并维持线粒体功能。HSP60在骨髓瘤细胞中的敲低（KD）导致增殖抑制并降低线粒体的质量。线粒体压力测试表明，HSP60 KD抑制糖酵解和线粒体活性。代谢组学显示糖酵解和三羧酸循环代谢物减少，并通过S反应抑制肌酸和磷酸肌酸的形成-腺苷甲硫氨酸（SAM），由去甲基腺苷转移酶1，线粒体（TFB1M）和减少的能量存储物质介导的氨基酸。此外，HSP60沉默通过磷酸戊糖途径影响了核糖核苷酸和烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的合成，从而抑制了细胞的增殖。HSP60 KD抑制5'腺苷单磷酸激活的蛋白激酶（AMPK），后者抑制关键酶6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3（PFKFB3），通过抑制丙二酰-丙二酸影响脂肪酸的代谢。辅酶A。我们的数据表明，HSP60表达降低会改变MM的代谢重编程，抑制肿瘤进程并减少线粒体依赖的生物合成，这表明HSP60是MM治疗的潜在治疗靶点。