Melnikova A K, Kuravsky M L, Kulikova K V, Sevostyanova I A, Muronetz V I
Faculty of Bioengineering and Bioinformatics Moscow State University, Moscow, Russia.
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia.
Biomed Khim. 2020 Sep;66(5):372-377. doi: 10.18097/PBMC20206605372.
The somatic isoform of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC1.2.1.12) is involved in such crucial for cancer cells development pathways as induction of apoptosis and glycolytic regulation. At the same time, sperm-specific isoform (GAPDHS) does not exhibit all the same functions as somatic enzyme. The expression of sperm-specific GAPDH without N-terminal domain in some melanoma cells along with somatic isoenzyme, shown in our previous work, has led to the proposal of this unusual enzyme's possible role in regulation of cancer cells glycolysis. In the presented work we have tested production of GAPDHS in 13 additional melanoma cell lines by immunoblotting. We have also gathered data on energy metabolism in 5 selected cell lines by evaluation of glucose uptake and lactate production in differing conditions. We have demonstrated that in standard cultivation media glucose uptake by MelP cells, producing substantial amounts of GAPDHS protein was higher than in MelKor cells, producing lesser amounts of GAPDHS. All other analyzed cell lines that do not produce GAPDHS (MelMS, MelSi and Malme3M) had even a lower glucose uptake rate.
糖酵解酶甘油醛-3-磷酸脱氢酶（GAPDH；EC1.2.1.12）的体细胞同工型参与了对癌细胞发育途径至关重要的过程，如诱导细胞凋亡和糖酵解调节。与此同时，精子特异性同工型（GAPDHS）并不具备与体细胞酶完全相同的功能。我们之前的研究表明，在一些黑色素瘤细胞中，不含N端结构域的精子特异性GAPDH与体细胞同工酶一同表达，这引发了关于这种特殊酶可能在癌细胞糖酵解调节中发挥作用的推测。在本研究中，我们通过免疫印迹法检测了另外13种黑色素瘤细胞系中GAPDHS的产生情况。我们还通过评估不同条件下的葡萄糖摄取和乳酸产生，收集了5种选定细胞系能量代谢的数据。我们证明，在标准培养基中，产生大量GAPDHS蛋白的MelP细胞的葡萄糖摄取量高于产生较少GAPDHS的MelKor细胞。所有其他不产生GAPDHS的分析细胞系（MelMS、MelSi和Malme3M）的葡萄糖摄取率甚至更低。
