ABSTRACT
Objective To investigate the tumor molecular mechanism of Hedgehog/Gli in promoting the epithelial-mesenchymal transition (EMT) in gastric cancer AZ521 cells. Methods After 24 h of treatment with GANT61,the mRNA expression of Gli1,Gli2, N-cadherin,and E-cadherin in the AZ521 cell line were detected by real-time fluorescence quantitative PCR. A Western blotting assay was conducted to determine the expression of the above cytokines,p-AKT and AKT. The effect of GANT61 on invasion was observed by transwell assay. N-Shh stimulation of the Hedgehog pathway was conducted to confirm the changes in these cytokines. Results GANT61 significantly downregulated the mRNA expression of Gli1,Gli2,and N-cadherin,but upregulated E-cadherin mRNA expression. The Western blotting assay revealed that GANT61 downregulated the protein expression of Gli1,Gli2,p-AKT,and N-cadherin,but upregulated E-cadherin expression. Furthermore,GANT61 inhibited the invasion. N-Shh proteins up-regulated Gli1,Gli2,and N-cadherin mRNA,protein expression and p-AKT protein expression,but downregulated E-cadherin mRNA and protein expressions. N-Shh promoted the invasion of tumor cells. Conclusion Downregulation of Gli1 and Gli2 can inhibit the invasion and metastasis in gastric cancer cells,which may be related to the promotion of EMT by Gli through the PI3K/AKT pathway.
ABSTRACT
Objective To study the inhibitory effects of GANT61, as an inhibitor of Gli, on the growth of human esophageal adenocarcinoma cell lines OE19 and OE33, and their mechanisms thereof. Methods After treating with different concentrations of GANT61(30,20,13.333 3,8.888 8,5.925 9,3.950 6,2.633 7,1.755 8,1.170 5μmol/L),the cell viabilities of OE19 and OE33 were detected by MTS method, which expressed by IC50. The Gli1and Gli2 mRNA expressions treated with GANT61(10 μmol/L GANT61) or DMSO for 24 h were detected in OE19 and OE33 cell lines by real time fluorescence quantitative PCR. The protein expressions of Gli1, Gli2 and CyclinD1 treated with GANT61 or DMSO for 24 h were detected in OE19 and OE33 cell lines by Western blot assay. Transwell invasion assay was performed to evaluate the inhibiting effect on OE19 and OE33 cell invasion by the treatment of GANT61 or DMSO. Results The IC50 of GANT61 was 8.08μmol/L in OE19 and 9.65μmol/L in OE33 cells. Compared with DMSO group, Gli1 and Gli2 mRNA expressions and Gli1,Gli2 and CyclinD1 protein expressions were significantly decreased in OE19 and OE33 cells of GANT61 group (P<0.05). The number of penetrating cells was significantly reduced in OE19 and OE33 cells of GANT61 group compared with that of DMSO group (P<0.01). Conclusion GANT61 can inhibit the growth and invasion of esophageal neoplasms cells by down-regulating Gli1 and Gli2 mRNA expression,which indicates that Hedgehog signaling pathway may play an important role in carcinogenesis and progression of esophageal adenocarcinoma.