The Role Of Cx43, Smad Mediated BMP And TGF-beta Signaling Pathways In The Occurrence And Development Of Gastriccarcinoma And The Relationship Among Them

Gastric carcinoma is one of common malignant tumour with highestincidence and mortality in our country. The occurrence of gastric carcinoma isa gradual process with various factors, stages and steps, which involves manysignal pathways that may arise intersection effects. Cx4plays a key role in cellsignal transmit. TGF-and Smads whose abnormal expresses are importantfactors on occurrence of gastric carcinoma are intercellular signal pathways.However, Smad1and Smad2are key component elements in transforminggrowth factor-which belongs to a super–family transmit pathway. Smad1and Smad2have possibility to be affected by intercellular transmit so thatgastric carcinoma occurs and develops.Comparing the expresses of Cx43, Smad1and Smad2in normal gastrictissue and the expresses in tissue of gastric carcinoma, analyzing relationshipbetween the three factors and clinical pathology index of gastric carcinoma(degree of differentiation, depth of invasion, lymph node metastasis etc.), thistrial offer theoretical basis to research gastric carcinoma, at the same timeoffer target spot to clinical treat.Part Ⅰ The relationship between Cx43,smad1, Smad2and gastriccarcinomaObjectiveTo detect the expression of Cx43,smad1and smad2in normal andcarcinomatous gastric tissue by immunohistochemisty, western blotting andRT-PCR, and reveal the relationship between them and gastric carcinomaoccurrence and development.MethodsTo collect60cases of gastric carcinoma and30cases of normal gastrictissue(tissue away the tumor more than10centimetres regarding as normal tissue for comparison) which were surgically removed and certified bypathology,and were not adopted by radiotherapy and chemotherapy inChengde Medical College affiliated hospital from September,2012to August,2013. Immunohistochemisty and western blotting were adopted to detect theexpression of Cx43,smad1and smad2in normal and gastric carcinoma tissue;the expression of Cx43mRNA,smad1mRNA and smad2mRNA was detectedby RT-PCR in normal and gastric carcinoma tissue. At the same time,analyzing the experimental data combined clinical pathology by statistics.Results1. Immunohistochemisty results1.1The expressions of Cx43，and smad1in carcinomatous gastric tissuewere lower than they were in normal gastric tissue (P<0.05). The expressionof smad2in carcinomatous gastric tissue was higher than that in normalgastric tissue (P<0.05)1.2The expressions of Cx43, smad1, smad2in gastric carcinoma tissuehave no statistical meaning with gender and age.(P＞0.05)1.3The expressions of Cx43and smad1in high differentiation groupwere higher than in low differentiation group(P<0.05), however, theexpressions of smad2was lower than that in poorly differentiation group(P<0.05).1.4The expression of Cx43and smad1in gastric carcinoma with lymphnode metastasis were lower than they were in gastric carcinoma withoutlymph node metastasis (P<0.05). The expression of smad2in gastriccarcinoma with lymph node metastasis was higher than that in gastriccarcinoma without lymph node metastasis (P<0.05).1.5The expressions of Cx43and smad1in gastric carcinoma with TNMⅢ+Ⅳ were lower than they were in gastric carcinoma with TNMⅠ+Ⅱ(P<0.05). The expression of smad2in gastric carcinoma was higher than thatin gastric carcinoma with TNMⅠ+Ⅱ (P<0.05). 2. western blotting detection2.1The expressions of Cx43，and smad1in carcinomatous gastric tissuewere lower than they were in normal gastric tissue (P<0.05). The expressionof smad2in carcinomatous gastric tissue was higher than that in normalgastric tissue (P<0.05)2.2The expressions of Cx43, smad1, smad2in gastric carcinoma tissuehave no statistical meaning with gender and age.(P＞0.05)2.3The expressions of Cx43and smad1in high differentiation groupwere higher than in low differentiation group(P<0.05), however, theexpressions of smad2was lower than that in poorly differentiation group(P<0.05).2.4The expressions of Cx43and smad1in gastric carcinoma withoutlymph node metastasis were higher than they were in gastric carcinoma withlymph node metastasis (P<0.05). The expression of smad2in gastriccarcinoma without lymph node metastasis was lower than that in gastriccarcinoma with lymph node metastasis (P<0.05).2.5The expressions of Cx43and smad1in gastric carcinoma with TNMⅢ+Ⅳ were lower than they were in gastric carcinoma with TNMⅠ+Ⅱ(P<0.05). The expression of smad2in gastric carcinoma was higher than thatin gastric carcinoma with TNMⅠ+Ⅱ (P<0.05).3. RT-PCR detection3.1The expressions of Cx43，and smad1mRNA in carcinomatous gastrictissue were lower than they were in normal gastric tissue (P<0.05). Theexpression of smad2mRNA in carcinomatous gastric tissue was higher thanthat in normal gastric tissue (P<0.05)3.2The expressions of Cx43mRNA, smad1mRNA, smad2mRNA ingastric carcinoma tissue have no statistical meaning with different gendergroups and different ages.(P＞0.05)3.3The expressions of Cx43mRNA and smad1mRNA in high andmedium differentiation group and were higher than in low differentiation group(P<0.05), however, the expressions of smad2mRNA was higher thanthat in poorly differentiation group (P<0.05).3.4The expressions of Cx43mRNA and smad1mRNA in gastriccarcinoma with lymph node metastasis were lower than they were in gastriccarcinoma without lymph node metastasis (P<0.05). The expression of smad2mRNA in gastric carcinoma with lymph node metastasis was higher than thatin gastric carcinoma without lymph node metastasis (P<0.05).3.5The expressions of Cx43mRNA and smad1mRNA in gastriccarcinoma with TNM Ⅲ+Ⅳ were lower than they were in gastric carcinomawith TNMⅠ+Ⅱ (P<0.05). The expression of smad2mRNA in gastriccarcinoma was higher than that in gastric carcinoma with TNMⅠ+Ⅱ(P<0.05).Conclusion1. The expressions of Cx43, smad1and smad2in gastric carcinomatous tissuehave marked differences from normal gastric tissue throughimmunohistochemistry check. And those expressions have closerelationships with clinical pathology index (degree of differentiation,lymph node transforming, TNM by stages), neither with age and gender.2. The expressions of Cx43, smad1and smad2in gastric carcinomatous tissuehave differences which have statistic meaning from normal gastric tissuethrough western blotting. And those expressions have relationships withclinical pathology index (degree of differentiation, lymph nodetransforming, TNM by stages), neither with age and gender.3. The expressions of Cx43mRNA, smad1mRNA and smad2mRNA ingastric carcinomatous tissue have differences which have statistic meaningfrom normal gastric tissue through RT-PCR check. And those expressionshave relationships with clinical pathology index (degree of differentiation,lymph node transforming, TNM by stages), neither with age and gender. Part Ⅱ The relationship among Cx43, smad1and smad2in the gastriccarcinomaObjectiveResearching the relationship among Cx43, smad1and smad2in carcinomabased on analyzing experimental data.MethodsAnalyzing experiment data of Cx43, smad1and smad2by statisticsoftware.Results1. From immunohistochemistry, the correlation coefficient Cx43andSmad1protein is:r=0.55(P<0.01);the correlation coefficient of Cx43andsmad2protein is:r=-0.52(P<0.01);the correlation coefficient smad1andsmad2protein is:r=-0.43(P<0.01).2. Through Western blotting, the correlation coefficient Cx43and Smad1protein is:r=0.67(P<0.01);the correlation coefficient of Cx43and smad2protein is:r=-0.67(P<0.01);the correlation coefficient smad1and smad2protein is:r=-0.65(P<0.01).3. Through RT-PCR, the correlation coefficient of Cx43mRNA andSmad1mRNA is:r=0.45(P<0.01);the correlation coefficient of Smad2mRNAand Cx43mRNA is:r=-0.53(P<0.01);the correlation coefficient of smad1mRNA and smad2mRNA is:r=-0.49(P<0.01).ConclusionsThe correlation of protein expression of Cx43and smad1was positive;the correlation of expression of Cx43mRNA and smad1mRNA was positive;the correlation of protein expression of Cx43and smad2was negative; thecorrelation of expression of Cx43mRNA and smad2mRNA was negative; thecorrelation of protein expression of smad1and smad2was negative. Thecorrelation of expression of smad1mRNA and smad2mRNA was negative.