Qu Yuan, Xue Shan, Zheng Yujian, Du Yajing, Zhang Guoping, Huang Liting, Li Hui, Li Huiwu
Department of Labour Hygiene and Sanitary Science, College of Public Health, Xinjiang Medical University, Urumqi, Xinjiang 830011, P.R. China.
Medical Research Center, Yuebei People's Hospital, Shantou University, Shaoguan, Guangdong 512025, P.R. China.
Oncol Lett. 2023 Aug 10;26(4):421. doi: 10.3892/ol.2023.14007. eCollection 2023 Oct.
Esophageal squamous cell carcinoma (ESCC) is an aggressive malignancy of the digestive system with increasing incidence and mortality rates. The biological roles of microRNA (miR)-378a-3p in tumor cells remain contested, and the mechanisms underlying the functions, energy metabolism, and cell survival mechanisms in ESCC cells are yet to be fully elucidated. In the present study, miR-378a-3p overexpression and negative control plasmids were transfected into ECA-109 cells using electroporation. Western blotting was used to detect the relative expression of proteins, and flow cytometry was used to detect cell apoptosis. Subsequently, ELISA assays were performed to determine enzyme activity, and an ATP detection kit was used to measure ATP content. Dual-luciferase reporter assays were performed to identify the target genes of miR-378a-3p. The results of the present study demonstrated that miR-378a-3p inhibited the gene expression and enzyme activities of glucose transporter protein 1 (GLUT-1), Aldolase A (ALDOA), and pyruvate kinase M2 (PKM2), all of which are involved in the glycolytic pathway of cells. Energy metabolism was suppressed by miR-378a-3p by reducing ATP content, and this downregulated the expression of Bcl-2 and Survivin. Moreover, increased miR-378a-3p expression promoted cell apoptosis in the early stages by increasing the expression levels and the activity of Bad and Caspase-3, while inhibiting the expression levels of Bcl-2 and Survivin. The results of the present study also demonstrated that GLUT-1/ALDOA/PKM2 were target genes of miR-378a-3p. Notably, miR-378a-3p blocked energy production and promoted the apoptosis of tumor cells via the downregulation of glycolytic enzyme expression and by reducing the mitochondrial membrane potential in ESCC. Bad, Caspase-3, Survivin, and Bcl-2 may be associated with blocking energy production and promoting apoptosis via miR-378a-3p in ESCC cells.
食管鳞状细胞癌（ESCC）是一种侵袭性消化系统恶性肿瘤，其发病率和死亡率呈上升趋势。微小RNA（miR）-378a-3p在肿瘤细胞中的生物学作用仍存在争议，ESCC细胞中其功能、能量代谢及细胞存活机制的潜在机制尚未完全阐明。在本研究中，采用电穿孔法将miR-378a-3p过表达质粒和阴性对照质粒转染至ECA-109细胞。使用蛋白质免疫印迹法检测蛋白质的相对表达，采用流式细胞术检测细胞凋亡。随后，进行酶联免疫吸附测定（ELISA）以确定酶活性，并使用ATP检测试剂盒测量ATP含量。进行双荧光素酶报告基因测定以鉴定miR-378a-3p的靶基因。本研究结果表明，miR-378a-3p抑制葡萄糖转运蛋白1（GLUT-1）、醛缩酶A（ALDOA）和丙酮酸激酶M2（PKM2）的基因表达及酶活性，这些蛋白均参与细胞的糖酵解途径。miR-378a-3p通过降低ATP含量抑制能量代谢，并下调Bcl-2和生存素的表达。此外, miR-378a-3p表达增加通过提高Bad和半胱天冬酶-3（Caspase-3）的表达水平及活性，同时抑制Bcl-2和生存素的表达水平，促进早期细胞凋亡。本研究结果还表明，GLUT-1/ALDOA/PKM2是miR-378a-3p的靶基因。值得注意的是，miR-378a-3p通过下调糖酵解酶表达并降低ESCC中的线粒体膜电位，阻断能量产生并促进肿瘤细胞凋亡。Bad、Caspase-3、生存素和Bcl-2可能与ESCC细胞中通过miR-378a-3p阻断能量产生和促进凋亡有关。
