The Effect Of Gli Inhibitor GANT61 On Esophageal Squamous Cell Carcinoma OE21

Objective: Esophagus cancer is one of the most common gastrointestinal tumor in our country and the morbidity and mortality rate is the highest in the world. The main pathological type of Esophagus is esophageal squamous cell carcinoma. At present, the effect of surgery, radiotherapy and chemotherapy to treat esophagus cancer is not satisfied. The prognosis of patients is very poor. More and more attention has been paid to the treatment of molecular targeted areas. The Hedgehog(Hh) signaling pathway plays a critical role during embryonic development in multicellular organisms. The activation of Hh signaling pathway involves in the regulation of adult stem cells in biological tissue, tissue damage repairment and regeneration of blood vessels and other physiological functions. As a final executor of Hh signaling pathway, continued aberrant activation of Gli causes the transcription of a variety of downstream cancer-related genes. Extensively study show that Gli abnormal activation occurs in a variety of human malignancies and Gli relates to the tumor occurrence and biological characteristics of malignant tumors. It has been discovered that increased abnormal expression of Gli in esophageal squamous biological tissue is closely relevant to the occurrence of tumor, and be considered as a independent factor of evaluation in survival prognosis. GANT6, a specific inhibitor of Hh signaling pathway transcription factor Gli, can induce tumor cells apoptosis of malignant mesothelioma, acute myeloid leukemia, rhabdomyosarcoma, pancreatic cancer and other tumors. Currently, there are very few investigations on GANT61 in human esophageal squamous. Here we mainly investigated the inhibition of esophageal squamous cell growth by GANT61, and the impact of GANT61 on esophageal squamous cell invasion and metastasis, explored the apoptosis mechanism of esophageal squamous cell and seek a low toxicity and efficient targeted drugs to treat esophageal squamous cancer.Methods: Using MTS method to test the viability inhibition of different concentrations of GANT61 treating 72 h of esophageal squamous carcinoma cell OE21(0,1.171,1.756,2.634,3.951,5.936,8.889,13.333,20,30μmol/L), cell viability curve drawing. Get calculation of IC50. Using Western blot method to detect the Gli1, Fas and cleaved caspase 3 protein expression of 10μmol/L GANT61 treating 24 h in esophageal squamous carcinoma cell OE21. Application of FCM to detect 10μmol/L GANT61 role 24 h affect esophageal squamous carcinoma cell OE21 apoptosis. Transwell invasion experiment testing 10μmol/L GANT61 treating 24 h effect on esophageal squamous carcinoma cell OE21 invasion ability.Result:1 The MTS experiment shows that the GANT61 is obvious cytotoxic to esophageal squamous carcinoma cell OE21, inhibit the tumor cell viability, with the increase of drug concentration, the cell viability present obvious downward trend. IC50=7.975μmol/L.2 According to the Western blot result, compared with control group, GANT61 can obviously decrease the Gli1 expression, increase the Fas and cleaved caspase 3 expression.3 The FCM result show that GANT61 treating 24 h in esophageal squamous carcinoma cell OE21 can induce cell apoptosis significantly, compared with control group, the difference was statistically significant(P<0.01).4 Transwell invasion experiment show that GANT61 treating 24 h in esophageal squamous carcinoma cell OE21 the number of cell migration decreased significantly, compared with control group, the difference was statistically significant(P<0.01).Conclusion: Gli1 inhibitor GANT61 is obvious cytotoxic to esophageal squamous carcinoma cell OE21, inhibits the tumor cell viability significantly, and has a dose dependent. IC50=7.975μmol/L. Western blot show that GANT61 can suppress Gli1 protein expression and increase cell apoptosis related protein Fas and cleaved caspase 3 expression, GANT61 may specific inhibit Gli1 protein expression in esophageal squamous carcinoma cell OE21 and decreases the Fas/FasL apoptotic cell death receptor pathway induce the apoptosis of esophageal squamous carcinoma cell. GANT61 can effectively restrain the invasion and metastasis ability of esophageal squamous carcinoma cell OE21. This study provide theoretical basis and new ideas for molecular targeted therapy of esophageal cancer.