The Regulation Of JAK2/STAT3 Signaling Pathway On Invasion And Metastasis Of Esophageal Cancer

Objective: To explore the effect of regulation of JAK2/STAT3 signaling pathway on tumor invasion and metastasis in esophageal squamous cell carcinoma,and to reveal the role of STAT3 signaling pathway in invasion and metastasis of esophageal squamous cell carcinoma.Methods: At the organizational level,179 cases of esophageal squamous cell carcinoma and adjacent tissues in the GEO database were used to statistically analyze the mRNA expression and survival prognosis of IL-6R,JAK2,STAT3 and PKM2.At the same time,IHC detected the relative expression of IL-6R and p-JAK2 protein in 100 cases of esophageal squamous cell carcinoma and corresponding 80 adjacent tissues in tissue microarray,and analyzed its correlation with clinicopathological features.At the cellular level,background expression of p-JAK2 and p-STAT3 was detected by western blot in seven esophageal squamous carcinoma cell lines,and KYSE150 esophageal scales with relatively high expression in p-JAK2 and p-STAT3 using lentiviral shRNA technology.The cancer cell line interfered with JAK2 and STAT3 by lentiviral infection,and the expression of p-JAK2 and p-STAT3 and the expression of STAT3 signaling pathway related factors were detected by western blot.MTT assay was used to detect the effects of cell proliferation,cell scratch assay and Transwell assay on cell migration and invasion,and flow cytometry assay on cell cycle and apoptosis rate.Results: At the organizational level: In the GEO database,the relative mRNA expression of JAK2 and PKM2 in esophageal squamous cell carcinoma was higher than that in adjacent tissues(P<0.0001,P<0.0001).The difference was statistically significant.IL-6R and STAT3 were in esophageal squamous There was no significant difference in the relative expression of mRNA between cancer and adjacent tissues(P=0.3514,P=0.5803).The relative mRNA expressions of IL-6R,JAK2,STAT3 and PKM2 were not correlated with the prognosis of patients(P=0.2910,P=0.1234,P=0.5025,P=0.2917);the Pearson correlation coefficient of IL-6R and PKM2 with STAT3(r = 0.347,r = 0.343)is higher than JAK2(r = 0.176).In the immunohistochemistry results of tissue microarray,the relative expression of IL-6R and p-JAK2 in esophageal squamous cell carcinoma was higher than that in adjacent tissues(P<0.001,P=0.002),but the difference was statistically significant;however,no correlation was found between IL-6R and p-JAK2 in the relative expression of protein in esophageal squamous cell carcinoma.The clinicopathological analysis showed that IL-6R and p-JAK2 protein were highly expressed in patients with gender(P=0.618,P=0.315).Age(P=0.122,P=0.446),tumor size(P=0.417,P=0.906),pathological grade(P=0.251,P=0.212),pathological morphology(P=0.103,P=0.699),infiltration depth(P=0.654,P=0.763),clinical stage(P=0.236,P=0.884),T stage(P=0.394,P=0.779),lymph node metastasis(P=0.436,P=0.656)were not statistically correlated.Kaplan-Meier survival analysis showed that there was no significant correlation between the relative expression of IL-6R and p-JAK2 protein and the prognosis of patients with esophageal squamous cell carcinoma(P=0.3754,P=0.9647).At the cellular level,Western Blot showed that JAK2 was significantly interfered with in esophageal cancer cell lines,p-JAK2 was significantly decreased,p-STAT3 was not found to be significantly inhibited,p-AKT,p-P70S6 K and p-PKM2 were increased;IL-6(50ng/ M))The drug was stimulated for 0.5 h,no significant change was found in p-JAK2,and p-AKT,p-P70S6 K,p-PKM2 and p-STAT3 were increased.Interfering with STAT3 in esophageal cancer cell lines,p-STAT3 was significantly decreased,p-JAK2,p-AKT,p-P70S6 K and p-PKM2 were not found to change significantly;IL-6(50ng/ml)drug stimulation 0.5h,p-STAT3 still has no obvious recovery.Cell function experiments showed that cell proliferation(P>0.05)and cell migration(P>0.05)were not significantly inhibited by JAK2;cell invasion was inhibited to some extent(P<0.05);cell cycle and apoptosis rate were not significantly changed.(P>0.05);interference with STAT3,cell proliferation(P<0.05),migration(P<0.05)and invasion(P<0.05)were significantly inhibited;cell cycle was also inhibited,and G0/G1 phase was shortened.The S phase was mainly prolonged,and the apoptosis rate was significantly increased(P<0.05).IL-6(50ng/ml)drug stimulation for 0.5h did not significantly affect cell function,cell cycle and apoptosis.Conclusion: 1.IL-6R,p-JAK2 and STAT3 may be involved in the malignant phenotype of esophageal cancer.2.Knockdown of STAT3 causes esophageal cancer cell cycle arrest in S phase,increasing apoptosis rate,inhibiting cell proliferation,migration and invasion.3.STAT3 is a key node of cell signaling pathway in esophageal cancer.In addition to JAK2 upstream,there may be other non-canonical(p-AKT/p-p70s6k/p-PKM2/p-STAT3)signaling pathways.