Glucose-6-Phosphate Dehydrogenase Promotes The Proliferation And Migration Of Lung Adenocarcinoma Cells Via The STAT3 Signaling Pathway

Objective: Lung cancer is the malignant tumor with the highest morbidity and mortality in China.Lung adenocarcinoma(LUAD)is the most common histopathological subtype.Glucose-6-phosphate dehydrogenase(G6PD)plays an important role in tumor progression by regulating the stability of intracellular dedox state and mediating nucleic acid synthesis.Signal transducers and activators of transcription3(STAT3)can activate the process of cell cycle and induction of invasion and metastasis related gene expression,which regulate the proliferation,differentiation and migration of tumor cells.This study explored the relationship between G6PD and lung adenocarcinoma,and observed how it affected the phenotypes of lung adenocarcinoma cells through the STAT3 signaling pathway.Methods: 1.60 fresh surgical specimens of lung adenocarcinoma were selected to observe the expression of G6PD in LUAD.We explored its correlation with clinicopathological characteristics and clinic prognostic significance.The online database was used to analyze the expression of G6PD in different LUAD subgroups and external databases were used to confirm its prognostic value.2.Firstly,RT-PCR and western blot(WB)methods were used to verify the expression of G6PD in different LUAC cell lines,and stable cell lines with G6PD gene interference and overexpression were constructed through lentivirus transfection.Subsequently,the experiment was divided into three groups: the G6PD interference group,the G6PD overexpression group and the negative control group.The effects of G6PD gene on the proliferation and migration of lung adenocarcinoma cells were observed by plate cloning formation,CCK-8,cell scar and transwell migration tests.In vivo experiments,the xenograft tumor model of nude mice was established,and then the size and quality of the xenograft tumor were measured,and the effect of overexpression of G6PD on the growth of xenograft tumor in lung adenocarcinoma was observed.3.The experiment was divided into four groups:NCI-H1299 cell line(NC group),H1299 cell line with G6PD overexpressed(OE group),H1299 cell line with G6PD overexpressed +STAT3 inhibitor(OE+Stattic group),NCI-H1299 cell line +STAT3 inhibitor(NC+Stattic group),negative control NCI-H1299 cell line +STAT3 inhibitor(NC+Stattic group).WB method was used to detect the expression of G6PD,STAT3,p STAT3,CyclinD1,p53 proteins in four groups.Then a rescue experiment(CCK-8,plate cloning formation,transwell migration tests)was designed to verify whether G6PD promotes the proliferation and migration of lung adenocarcinoma cells through the STAT3 signaling pathway.Results: 1.The high expression of G6PD was significantly correlated with the tumor size,TNM stage and lymph node metastasis of lung adenocarcinoma.Compared with patients with low expression of G6PD,the postoperative survival time of patients with high expression of G6PD was significantly shortened(log-rank test,p < 0.001).The online database shows that the expression level of G6PD in lung adenocarcinoma tissues varies to different degrees in terms of gender,smoking status,and stage,and the high expression of G6PD may indicate a shorter survival time.2.The H1299 cell lines with the highest G6PD expression were successfully screened for follow-up experiments.Stable transfected cell lines with G6PD interference and overexpression were successfully constructed.In vitro experiments,overexpression of G6PD can promote the proliferation and migration of lung adenocarcinoma cells,while interference with the expression of G6PD is the opposite.In vivo experiments in nude mice,compared with nude mice transplanted with normal lung adenocarcinoma cells expressing G6PD gene,the transplanted lung adenocarcinoma cells with overexpression of G6PD showed stronger proliferation ability.3.In lung adenocarcinoma cells,the overexpression of G6PD can significantly promote the activation of the STAT3 signaling pathway,while the expression of CyclinD1 is up-regulated and the expression of p53 is inhibited.G6PD promotes the proliferation and migration of lung adenocarcinoma cells through the STAT3 signaling pathway.Conclusion: 1.The high expression of G6PD is closely related to the prognosis of patients with lung adenocarcinoma,which can be used as a tumor marker to effectively evaluate the prognosis and survival of patients with lung adenocarcinoma.2.Overexpression of G6PD can promote the proliferation of lung adenocarcinoma cells and increase their movement and migration ability,while interference with G6PD expression is the opposite.3.The overexpression of G6PD can significantly promote the activation of the STAT3 signaling pathway of lung adenocarcinoma cells,promote CyclinD1 and inhibit the expression of p53.G6PD increases the proliferation and migration of lung adenocarcinoma cells through the STAT3 signaling pathway.