Department of Experimental and Clinical Biomedical Sciences, University of Florence, Viale G.B. Morgagni 50, 50134 Firenze, Italy.
Department of Experimental and Clinical Medicine, University of Florence, Largo Brambilla 3, 50134 Firenze, Italy.
Cells. 2020 Jan 28;9(2):308. doi: 10.3390/cells9020308.
Urokinase Plasminogen Activator (uPA) Receptor (uPAR) is a well-known GPI-anchored three-domain membrane protein with pro-tumor roles largely shown in all the malignant tumors where it is over-expressed. Here we have exploited the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 gene knock out approach to investigate its role in the oxidative metabolism in human melanoma and colon cancer as the consequences of its irreversible loss. Knocking out , a uPAR-encoding gene, in A375p, A375M6 and HCT116, which are two human melanoma and a colon carcinoma, respectively, we have observed an increased number of mitochondria in the two melanoma cell lines, while we evidenced an immature biogenesis of mitochondria in the colon carcinoma culture. Such biological diversity is, however, reflected in a significant enhancement of the mitochondrial spare respiratory capacity, fueled by an increased expression of GLS2, and in a decreased glycolysis paired with an increased secretion of lactate by all uPAR KO cells. We speculated that this discrepancy might be explained by an impaired ratio between LDHA and LDHB.
尿激酶型纤溶酶原激活物受体 (uPAR) 是一种众所周知的 GPI 锚定三结构域膜蛋白，其在所有过度表达的恶性肿瘤中发挥着促肿瘤作用。在这里，我们利用规律成簇间隔短回文重复序列 (CRISPR)/Cas9 基因敲除方法来研究其在人类黑色素瘤和结肠癌的氧化代谢中的作用，这是其不可逆丢失的结果。在两种人黑色素瘤细胞系 A375p 和 A375M6 以及结肠癌细胞系 HCT116 中敲除编码 uPAR 的基因 ，我们观察到这两种黑色素瘤细胞系中的线粒体数量增加，而在结肠癌细胞系中则存在未成熟的线粒体生物发生。然而，这种生物学多样性反映在由 GLS2 表达增加驱动的线粒体备用呼吸能力的显著增强，以及伴随着所有 uPAR KO 细胞中乳酸盐分泌增加的糖酵解减少。我们推测，这种差异可能是由 LDHA 和 LDHB 之间的比例失调引起的。
