Chair of Medical Biochemistry, Faculty of Medicine, Jagiellonian University Medical College, Kopernika 7 St., 31-034 Krakow, Poland.
Biomolecules. 2023 Oct 7;13(10):1491. doi: 10.3390/biom13101491.
This study was performed on human primary (WM115) and metastatic (WM266-4) melanoma cell lines developed from the same individual. The expression of proteins involved in L-cysteine metabolism (sulfurtransferases, and cystathionine β-synthase) and antioxidative processes (thioredoxin, thioredoxin reductase-1, glutathione peroxidase, superoxide dismutase 1) as well as the level of sufane sulfur, and cell proliferation under hypoxic conditions were investigated. Hypoxia in WM115 and WM266-4 cells was confirmed by induced expression of carbonic anhydrase IX and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 4 by the RT-PCR and Western blot methods. It was shown that, under hypoxic conditions the inhibition of WM115 and WM266-4 melanoma cell proliferation was associated with decreased expression of thioredoxin reductase-1 and cystathionine β-synthase. These two enzymes may be important therapeutic targets in the treatment of melanoma. Interestingly, it was also found that in normoxia the expression and activity of 3-mercaptopyruvate sulfurtransferase in metastatic WM266-4 melanoma cells was significantly higher than in primary melanoma WM115 cells.
本研究以源自同一患者的人原发性(WM115)和转移性(WM266-4)黑素瘤细胞系为研究对象。研究了参与 L-半胱氨酸代谢(硫转移酶和胱硫醚β-合酶)和抗氧化过程(硫氧还蛋白、硫氧还蛋白还原酶 1、谷胱甘肽过氧化物酶、超氧化物歧化酶 1)的蛋白质表达水平,以及 suffane 硫水平和缺氧条件下的细胞增殖情况。通过 RT-PCR 和 Western blot 方法证实了 WM115 和 WM266-4 细胞中的缺氧,即碳酸酐酶 IX 和 6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 4 的诱导表达。结果表明,在缺氧条件下,WM115 和 WM266-4 黑素瘤细胞增殖的抑制与硫氧还蛋白还原酶 1 和胱硫醚β-合酶表达的降低有关。这两种酶可能是治疗黑素瘤的重要治疗靶点。有趣的是,还发现,在常氧条件下,转移性 WM266-4 黑素瘤细胞中 3-巯基丙酮酸硫转移酶的表达和活性明显高于原发性黑素瘤 WM115 细胞。