The Molecular Mechanism Of Sonic Hedgehog In Squamous Cell Lung Cancer

Background:Lung squamous cell carcinoma is one of the most common tumors in China.The incidence rate in recent years has been increasing,which seriously threatens human health.There are still many mysteries in the pathogenesis and development of lung cancer.The constant revealing of these mysteries will provide more targets for individualized treatment of lung cancer.Hedgehog(Hh)signaling pathway has important influence on the normal development process such as fetal lung morphogenesis and function establishment,and it is indispensable for lung morphogenesis.A number of studies on the role of the Hh signaling pathway(also known as the Shh signaling pathway)in tumors have revealed that they include various types of tumors including human lung squamous cell carcinoma,gastric cancer,pancreatic cancer,basal basal cell carcinoma of the skin,ovarian cancer,and prostate cancer.The occurrence and development of the abnormal activation.At present,more and more research evidence shows that Hh signaling pathway is closely related to squamous cell carcinoma of the lung.Immunohistochemistry shows that the members of the signaling pathway are highly expressed in the tissues of lung squamous cell carcinoma patients.The GATA transcription factor contains an evolutionarily conserved C2 zinc finger DNA domain.In recent years,GATA6,one of the GATA family,has become more and more in-depth.According to reports,GATA6 regulates Hh signaling pathway activity,conditionally deleting GATA6 leads to ectopic expression of Shh and its transcriptional targets.However,the mechanism of the high expression of Shh in lung squamous cell carcinoma and the role of Shh in lung squamous cell carcinoma are not yet clear.This study will study the role of Shh on the migration and invasion of lung squamous cell carcinoma cells and elucidate the molecular mechanisms of Shh enrichment in lung cancer,thus providing a theoretical basis for the prognosis and treatment of lung cancer.Objectives:1.To explore the molecular mechanism of high expression of Shh ligand in lungsquamous cell carcinoma and to clarify the role of Shh in the pathogenesis of lung squamous cell carcinoma;2.To reveal the role of regulating Shh levels in the migration and invasion of lung squamous cell carcinoma and to provide new interventions for clinical treatment of lung squamous cell carcinoma Methods:1.Clinical tissue specimen detection1)Collect pathological diagnosis data and screen 111 cases of lung squamous cell carcinoma patients without other diseases and without radiotherapy and chemotherapy before surgery.2)The collected specimens were fixed and the content of Shh was detected by immunohistochemistry.2.Exogenous N-Shh stimulated lung squamous cell carcinoma,and the invasion and migration of lung squamous carcinoma cells were observed by Transwell and scratch experiments.3.Using shuffle virus to interfere or over-express Shh in SK-MES-1 and H226 cells,and to observe the invasion and migration of lung squamous cell carcinoma cells by Transwell and scratch experiments.4.Bioinformatics analysis of GATA6 expression in lung squamous cell carcinoma,Western blot and qPCR assay GATA6 on the regulation of Shh expression,Transwell assay Gat6-induced Shh-induced lung squamous cell carcinoma invasion and migration.5.Dual-luciferin reporter gene and chromatin immunoprecipitation experiments explore the mechanism of GATA6 regulating Shh expression.Results:1.The detection results of clinical tissue specimens showed that Shh ligand is highly expressed in lung squamous cell carcinoma tissues,and the level of Shh isinversely proportional to the prognosis of lung squamous cell carcinoma patients.2.Transwell results showed that exogenous N-Shh promoted the invasion and migration of lung squamous carcinoma cells.3.Transwell assay results showed that the high expression of Shh promoted the invasion and migration of lung squamous cell carcinoma cells,and interfered with Shh to decrease the invasion and migration of lung squamous cell carcinoma cells.4.GATA6 low expression in lung squamous cell carcinoma,GATA6 affect Shh expression levels decreased and cell migration and invasion decreased5.The results of dual-fluorescein reporter gene and chromatin immunoprecipitation experiments show that GATA6 is an inhibitory transcription factor regulating Shh expression.Conclusion:The expression of Shh in lung squamous cell carcinoma is high and theexpression of GATA6 is low.GATA6 is a Shh-inhibitory transcription factor.GATA6 inhibits the expression of Shh and thus affects the invasion and migration of lung squamous cell carcinoma cells.