Silencing of ENO1 by shRNA Inhibits the Proliferation of Gastric Cancer Cells

Abstract

α-Enolase is a significant subunit of enolase and acts as a glycolytic enzyme responsible for catalyzing the conversion of 2-phosphoglycerate to phosphoenolpyruvate in the anaerobic glycolysis pathway. The research about their role is known little in tumor invasion and metastasis. This research analyzed the effect of α-enolase in proliferation and progression of human gastric cancer. The constructed PLKO.1-ENO1 shRNA vector was transfected into 293 T cells and used to infect gastric cancer cells, MKN45, by using lentivirus method. Negative controls were generated by infection with viruses containing empty vector PLKO.1-scramble-shRNA by the same protocol and using wild-type MKN45 cells as blank control. The silencing effect was confirmed by reverse transcription polymerase chain reaction and Western blotting at messenger RNA and protein levels, respectively. Cell proliferation and chemosensitivity were tested by methyl-thiazolyl-tetrazolium assay. Cell apoptosis was tested by flow cytometry. The cell line α-enolase short hairpin RNA stabling silence α-enolase was successfully constructed. In the α-enolase short hairpin RNA cell lines, messenger RNA and protein expression of α-enolase were significantly lower than those in negative control and blank control groups. The proliferation and clone formation ability were significantly inhibited, cell apoptosis was increased significantly, and the inhibition rate of chemotherapy drugs was increased (P < .05). Our data provide strong evidence that α-enolase short hairpin RNA interference vector can effectively suppress the proliferation and increase chemosensitivity of MKN45 cells, which may provide a novel gene therapy for gastric cancer.

Enolase 1 Regulates Stem Cell-Like Properties in Gastric Cancer Cells by Stimulating Glycolysis

“…Recently, It has been shown that ENO1 expression is abnormal in many human cancers, including glioma, colorectal cancer, pancreatic cancer, lung cancer, and head and neck cancers [28,29,31,45,46]. Furthermore, previous studies have demonstrated that ENO1 was overexpressed in GC tissues and was related to the progression and prognosis of GC [35,36]. In this study, we further demonstrated that ENO1 expression was signi cantly associated with the overall survival of GC patients, implying the important functions of ENO1 in GC progression.…”

“…For example, ENO1 is related to the proliferation and metastasis of GCs [35]. Additionally, overexpression of ENO1 can promote cisplatin resistance by enhancing glycolysis in GCs, while in contrast, inhibition of ENO1 can increase the sensitivity of GCs to chemotherapy by repressing glycolysis [36].…”

Metabolic Pathways Enhancement Confers Poor Prognosis in p53 Exon Mutant Hepatocellular Carcinoma

“…19 ENO1 silencing by shRNA has been shown to effectively suppress the proliferation and increase chemosensitivity of gastric cells. 20 We proposed big data sets collected from TCGA to obtain complete gene expression profiles in p53 status. The GSEA method is responsible for 11 799 genes expression of an entire database evaluation that can execute the calculation of an ES, estimate the significance level of ES, and adjust for multiple hypothesis testing.…”

Unlikely role of glycolytic enzyme ��-enolase in cancer metastasis and its potential as a prognostic biomarker

“…Conversely, silencing of a-enolase in glioma, pancreatic, lung, endometrial, colorectal and breast cancer cells was found to induce cell cycle arrest and senescence, and also to reduce tumour volume in CFPAC-1 pancreatic, MDA-MB-231 breast and U-87MG glioma xenograft models in vivo [6,11,12,62,63] . Furthermore, a-enolase is also implicated in the control of apoptosis and sensitivity to chemotherapeutic agents, as silencing of ENO1 in cancer cells induced apoptosis and increased sensitivity to cisplatin and 5-fluorouracil in vitro [13,62,64] . Unexpectedly, cells respond to a-enolase silencing by inducing catabolic adaptations that lead to restoration of pyruvate, acetyl-CoA bulk and oxidative phosphorylation, and exhibit an increased expression of proteins involved in both oxidative stress-and sirtuin-induced autophagy [62] .…”

“…Overexpression promoted cell proliferation and tumour growth in vivo ; Decreased expression decreased cell proliferation and tumour growth in vivo [11] Endometrial carcinoma Knockdown in HEC-1B and Ishikawa Decreased expression reduced cell proliferation in vitro and tumourigenesis in vivo [12] Gastric cancer Knockdown in MGC-803 and MKN45 cells Knockdown led to cell cycle arrest at the G 1 phase and promoted apoptosis, and repressed the rate of cell proliferation and colony formation [13] Knockdown in MKN45 cells Knockdown decreased cell proliferation, induced apoptosis and increased sensitivity to chemotherapeutics [64]…”