The Expression Of ADD1 And Its Relationship With Hedgehog Signaling Pathway In Gastric Cancer

Objective:To detect the expression of ADD1 and to explore its relationship with Hedgehog signaling pathway in gastric cancer tissue,as well as providing a new method to clear the development mechanism of Gastric cancer.Methods:We had collected 60 cases of normal tissue and 60 caces of gastric cancer tissue in Chengde Medical College Affiliated Hospital from September 2013 to June 2015,these collected patients had not taken radiotherapy,chemotherapy and was diagnosed with gastric cancer by pathological examination after operation.We had used three methods included Western blotting,Immunohistochemistry,QReal-Time PCR technology to detected the expression of ADD1,IHH,PTCH1,GLI1 protein and mRNA in normal tissue and gastric cancer tissue.Then we analyzed the experimental data combined with clinical data,the difference was statistically significant when P<0.05.Results:1.The results of Western Blot1.1 Protein expression of ADD1,IHH,PTCH1 and GLI1 in normal and gastric carcinoma tissuesThe relative expression levels of ADD1,IHH,PTCH1,and GLI1 protein in gastric cancer tissues were higher than normal gastric tissues(P<0.05).1.2 The connection between clinicopathological features and the protein expression of ADD1,IHH,PTCH1,GLI1 in gastric carcinomaIn gastric carcinoma the protein expression of ADD1,IHH,PTCH1 and GLI1 was not significantly correlated with age and sex of patients(P>0.05).The protein expression levels of ADD1,IHH,PTCH1 and GLI1 in poorly differentiated gastric cancer tissues were higher well differentiated gastric carcinoma(P<0.05)In Ⅰ and Ⅱ stage of TNM the protein expression levels of ADD1,IHH,PTCH1 and GLI1 were lower than those Ⅱ Ⅰ and IV stage of TNM in gastric cancer(P<0.05).The relative protein expression levels of ADD1,IHH,PTCH1 and GLI1 with serosal invasion were higher than those with no serosal invasion in gastric carcinoma tissues(P<0.05).The relative protein expression levels of ADD1,IHH,PTCH1 and GLI1 with lymph node metastasis were higher than those with no lymph node metastasisin gastric cancer tissues(P<0.05).2.Results of Immunohistochemistry2.1 Protein expression of ADD1,IHH,PTCH1 and GLI1 in normal and gastric carcinoma tissuesThe relative expression levels of ADD1,IHH,PTCH1,and GLI1 protein in gastric cancer tissues were higher than normal gastric tissues(P<0.05).2.2 The connection between clinicopathological features and the protein expression of ADD1,IHH,PTCH1,GLI1 in gastric carcinomaIn gastric carcinoma the protein expression of ADD1,IHH,PTCH1 and GLI1 was not significantly correlated with age and sex of patients(P>0.05).The protein expression levels of ADD1,IHH,PTCH1 and GLI1 in poorly differentiated gastric cancer tissues were higher than well differentiated gastric carcinoma(P<0.05).In Ⅰ and Ⅱ stage of TNM the protein expression levels of ADD1,IHH,PTCH1 and GLI1 were lower than those Ⅱ Ⅰ and IV stage of TNM in gastric cancer(P<0.05).The relative protein expression levels of ADD1,IHH,PTCH1 and GLI1 with serosal invasion were higher than those with no serosal invasion in gastric carcinoma tissues(P<0.05).The relative protein expression levels of ADD1,IHH,PTCH1 and GLI1 with lymph node metastasis were higher than those with no lymph node metastasisin gastric cancer tissues(P<0.05).3.Results of QReal-Time PCR3.1 The mRNA expression of ADD1,IHH,PTCH1 and GLI1 in normal and gastric carcinoma tissuesThe relative mRNA expression levels of ADD1,IHH,PTCH1,and GLI1 in gastric cancer tissues were higher than normal gastric tissues(P<0.05).3.2 The relationship between mRNA expression of ADD1,IHH,PTCH1,GLI1 and clinicopathological features in gastric carcinomaIn gastric carcinoma the mRNA expression of ADD1,IHH,PTCH1 and GLI1 was not significantly correlated with age and sex of patients(P>0.05).The mRNA expression levels of ADD1,IHH,PTCH1 and GLI1 in poorly differentiated gastric cancer tissues were higher than well differentiated gastric carcinoma(P<0.05).In Ⅰ and Ⅱ stage of TNM the mRNA expression levels of ADD1,IHH,PTCH1 and GLI1 were lower than those Ⅱ Ⅰ and IV stage of TNM in gastric cancer(P<0.05).The relative mRNA expression levels of ADD1,IHH,PTCH1 and GLI1 with serosal invasion were higher than those with no serosal invasion in gastric carcinoma tissues(P<0.05).The relative mRNA expression levels of ADD1,IHH,PTCH1 and GLI1 with lymph node metastasis were higher than those with no lymph node metastasisin gastric cancer tissues(P<0.05).4.Correlation of ADD1,IHH,PTCH1 and GLI1 in gastric carcinomaCorrelation analysis of Western Blotting data:the relative protein expression of ADD1 correlated positively with the relative protein expression of IHH(r=0.611,P<0.05);the relative protein expression of ADD1 correlated positively with the relative protein expression of PTCH1(r=0.681,P<0.05);the relative protein expression of ADD1 correlated positively with the relative protein expression of GLI1(r=0.759,P<0.05).Correlation analysis of Immune histochemical data:the relative protein expression of ADD1 correlated positively with the relative protein expression of IHH(r=0.532,P<0.05);the relative protein expression of ADD1 correlated positively with the relative protein expression of PTCH1(r=0.731,P<0.05);the relative protein expression of ADD1 correlated positively with the relative protein expression of GLI1(r=0.852,P<0.05).Correlation analysis of QRealTime-PCR data:the relative mRNA expression of ADD1 correlated positively with the relative mRNA expression of IHH:(r=0.675,P<0.05);the relative mRNA expression of ADD1 correlated positively with the relative mRNA expression of PTCH1(r=0.811,P<0.05);the relative mRNA expression of ADD1 correlated positively with the relative mRNA expression of GLI1(r =0.719,P<0.05).Conclusion:1.The expression level of ADD1 in gastric cancer tissues was higher than that in normal gastric tissues,which was significantly higher than that of well differentiated gastric cancer tissues in poorly differentiated gastric cancer tissues.Therefore,which suggested that ADD1 plays an important role in the occurrence and development of gastric cancer.2.In gastric cancer tissues the expression levels of ADD1 with serosal invasion were higher than tissues with no serosal invasion,the expression levels of ADD1 in gastric cancer tissues with lymph node metastasis were higher than tissues with no lymph node metastasisin.These results suggested that the high expression of ADD1 can promote the invasion and metastasis of gastric cancer.3.The expression level of ADD1 is positively correlated with the expression of Hedgehog signaling pathway IHH,PTCH1,GLI1,so we speculate that there is a relationship between ADD1 and Hedgehog signaling pathway in the occurrence and development of gastric cancer.