Long Niya, Peng Shuo, Chu Liangzhao, Jia Jun, Dong Minghao, Liu Jian
Department of Pathology, Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China.
Key Laboratory of Endemic and Ethnic Diseases, (Guizhou Medical University) Ministry of Education, Guiyang, Guizhou 550004, P.R. China.
Exp Ther Med. 2020 Nov;20(5):72. doi: 10.3892/etm.2020.9200. Epub 2020 Sep 9.
Energy metabolic reprogramming (EMR) allows for the rearrangement of a series of metabolic genes and proteins when tumor cells adapt to their microenvironment. EMR is characterized by changes in the metabolic pattern and metabolic intermediates to meet the needs of tumor cells for their malignant proliferation and infiltrative growth. The present study investigated the role of low-dose paclitaxel (PTX) in changing the expression levels of key genes and proteins during glycolysis in CD133+ U251 glioma cells and explored the relevant regulatory mechanisms of action at the molecular level. CD133 immunomagnetic beads were applied to malignant CD133+ U251 glioma cells, which were then divided into a negative control and an experimental group treated with 1, 2, 4 or 8 µM PTX for 72 h. Cell Counting Kit-8 (CCK-8) was used to measure U251 cell proliferation. RNA and protein were extracted from the malignant glioma cells in all groups to observe changes in the expression levels of key glycolytic enzymes, such as glucose transporter 1 (GLUT1), pyruvate kinase M (PKM) and lactate dehydrogenase A (LDHA), using reverse transcription-quantitative PCR and western blot assays. Transwell migration assays were performed to quantify the effects of PTX solution on U251 cells. CD133+ U251 glioma cells were isolated successfully. CD1133+ cells had a higher rate of proliferation compared with CD1133- cells. In CD1133+ cells treated with PTX, a dose-dependent reduction in the expression levels of the key glycolytic enzymes GLUT1, PKM and LDHA was observed at both the mRNA and protein levels. PTX solution also inhibited cell migration. Differences between the control and experimental groups were statistically significant (P<0.05). Since glycolysis plays an indispensable role in the proliferation and migration of stem cell-like glioma cells, PTX may inhibit tumor cell growth by downregulating the gene and protein expression levels of glycolytic enzymes in CD133+ glioma cells.
能量代谢重编程（EMR）使肿瘤细胞在适应其微环境时能够重新排列一系列代谢基因和蛋白质。EMR的特征在于代谢模式和代谢中间体的变化，以满足肿瘤细胞恶性增殖和浸润性生长的需求。本研究调查了低剂量紫杉醇（PTX）在改变CD133 + U251胶质瘤细胞糖酵解过程中关键基因和蛋白质表达水平方面的作用，并在分子水平上探索了相关的作用调节机制。将CD133免疫磁珠应用于恶性CD133 + U251胶质瘤细胞，然后将其分为阴性对照组和用1、2、4或8μM PTX处理72小时的实验组。使用细胞计数试剂盒-8（CCK-8）测量U251细胞增殖。从所有组的恶性胶质瘤细胞中提取RNA和蛋白质，使用逆转录定量PCR和蛋白质印迹分析观察关键糖酵解酶（如葡萄糖转运蛋白1（GLUT1）、丙酮酸激酶M（PKM）和乳酸脱氢酶A（LDHA））表达水平的变化。进行Transwell迁移试验以量化PTX溶液对U251细胞的影响。成功分离出CD133 + U251胶质瘤细胞。与CD1133-细胞相比，CD1133 +细胞具有更高的增殖率。在用PTX处理的CD1133 +细胞中，在mRNA和蛋白质水平上均观察到关键糖酵解酶GLUT1、PKM和LDHA表达水平呈剂量依赖性降低。PTX溶液也抑制细胞迁移。对照组和实验组之间的差异具有统计学意义（P<0.05）。由于糖酵解在干细胞样胶质瘤细胞的增殖和迁移中起着不可或缺的作用，PTX可能通过下调CD133 +胶质瘤细胞中糖酵解酶的基因和蛋白质表达水平来抑制肿瘤细胞生长。
