The Expressions And Significance Of Cx43and PI3k/Akt Signaling Pathway In The Occurrence And Development Of Gastric Carcinoma And The Relationship Among Them

Gastric cancer is one of the malignant tumors with a high morbidity andmortality around the world, becoming a great threat to human health. Atpresent the pathogenesis of gastric cancer is not completely known, itsoccurrence and development is a biology process that has many stages, andsteps, it closely associated with abnormal changes of tumor cell signaltransduction. Gap junctions are cell membrane protein channels composed ofconnexin proteins that allow signal transduction between adjacent cells. It isthe structural basis of gap junctional intercellular communication. A lot of datasuggest that aberrant gap junction often involves in variety of cancer models,it relatives with tumorgenesis and metastasis. Cx43is the most widely studiedconnexin nowadays.PI3k/Akt signaling pathway is one of the importantsignaling pathways inside the cell, its activation can induce anti-apoptosisproliferation and angiogenesis and plays an essential role in the genesis anddevelopment of tumor. To preliminary study the relationship between gastriccancer intracellular and intercellular signaling, we detected the expression ofCx43,PI3Kand Akt in gastric cancer and normal control groups, observed therelationship between Cx43and PI3k/Akt in gastric cancer, to explore thepathogenesis of gastric cancer and try to find a new way of therapy in thisdisease.Part Ⅰ The relationship between Cx43,PI3K, Akt and gastriccarcinomaObjectiveTo detect the expression of Cx43, PI3K and Akt in 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 and25cases of normal gastric tissue(tissue away the tumor more than10centimetres regarding as normaltissue for comparsion) which were surgically removed and certified bypathology, and were not adopted by radiotherapy and chemotherapy inChengde Medical College affiliated hospital from July2012to February2013.Immunohistochemisty and western blotting were adopted to detect theexpression of Cx43, PI3K and Akt in normal and carcinomatous gastric tissue;the expression of Cx43mRNA, PI3KmRNA and AktmRNA was detected byRT-PCR in normal and carcinomatous gastric tissue. To analyse theexperimental data we combined clinical pothology with statistics.Results1. Immunohistochemisty results1.1The expression of Cx43, PI3K and Akt in different tissue1.1.1The expression of Cx43in carcinomatous gastric tissue were lowerthan they were in normal gastric tissue (P<0.05). The expression of PI3K andAkt in carcinomatous gastric tissue was higher than that in normal gastrictissue (P<0.05).1.2The expression of Cx43, PI3K and Akt in different age and genders1.2.1The expression of Cx43, PI3K and Akt in group of male and femaleand group of＞56and≤56years is not significant.1.3The expression of Cx43, PI3K and Akt in different clinicopathologicfeatures of carcinomatous gastric tissue1.3.1The expression of Cx43in gastric carcinoma which were welldifferentiated and moderately differentiated were higher than they were inpoorly differentiated gastric carcinoma (P<0.05).The expression of PI3K andAkt in gastric carcinoma which were well differentiated and moderatelydifferentiated was lower than that in poorly differentiated gastric carcinoma(P<0.05).1.3.2The expression of Cx43in gastric carcinoma without lymph nodemetastasis were higher than they were in gastric carcinoma with lymph nodemetastasis (P<0.05). The expression of PI3K and Akt in gastric carcinomawithout lymph node metastasis was lower than that in gastric carcinoma with lymph node metastasis (P<0.05).1.3.3The expression of Cx43in gastric carcinoma with TNM Ⅰ/Ⅱ werehigher than they were in gastric carcinoma with TNM Ⅲ/Ⅳ (P<0.05). Theexpression of PI3K and Akt in gastric carcinoma with TNM Ⅰ/Ⅱ was lowerthan that in gastric carcinoma with TNM Ⅲ/Ⅳ (P<0.05).1.3.4The expression of Cx43in invasion of gastric carcinoma not gettingin touch with serosa were higher than they were in gastric carcinoma withgetting in touch with serosa (P<0.05). The expression of PI3K and Akt ininvasion of gastric carcinoma not getting in touch with serosa was lower thanthat in gastric carcinoma with getting in touch with serosa (P<0.05).2. western blotting detection2.1The expression of Cx43, PI3K and Akt protein in different tissue2.1.1The expression of Cx43in carcinomatous gastric tissue were lowerthan they were in normal gastric tissue (P<0.05). The expression of PI3K andAkt in carcinomatous gastric tissue was higher than that in normal gastrictissue (P<0.05)2.2The expression of Cx43, PI3K and Akt protein in different age andgenders2.2.1The expression of Cx43, PI3K and Akt in group of male and femaleand group of＞56and≤56years is not significant.2.3The expression of Cx43, PI3K and Akt protein in differentclinicopathologic features of carcinomatous gastric tissue2.3.1The expression of Cx43in gastric carcinoma which were welldifferentiated and moderately differentiated were higher than they were inpoorly differentiated gastric carcinoma (P<0.05).The expression of PI3K andAkt in gastric carcinoma which were well differentiated and moderatelydifferentiated was lower than that in poorly differentiated gastric carcinoma(P<0.05).2.3.2The expression of Cx43in gastric carcinoma without lymph nodemetastasis were higher than they were in gastric carcinoma with lymph nodemetastasis (P<0.05). The expression of PI3K and Akt in gastric carcinoma without lymph node metastasis was lower than that in gastric carcinoma withlymph node metastasis (P<0.05).2.3.3The expression of Cx43in gastric carcinoma with TNM Ⅰ/Ⅱ werehigher than they were in gastric carcinoma with TNM Ⅲ/Ⅳ (P<0.05). Theexpression of PI3K and Akt in gastric carcinoma with TNM Ⅰ/Ⅱ was lowerthan that in gastric carcinoma with TNM Ⅲ/Ⅳ (P<0.05).2.3.4The expression of Cx43in invasion of gastric carcinoma not gettingin touch with serosa were higher than they were in gastric carcinoma withgetting in touch with serosa (P<0.05). The expression of PI3K and Akt ininvasion of gastric carcinoma not getting in touch with serosa was lower thanthat in gastric carcinoma with getting in touch with serosa (P<0.05).3. RT-PCR detection3.1The expression of Cx43, PI3K and Akt mRNA in different tissue3.1.1The expression of Cx43mRNA in carcinomatous gastric tissuewere lower than they were in normal gastric tissue (P<0.05). The expressionof PI3K and Akt mRNA in carcinomatous gastric tissue was higher than thatin normal gastric tissue (P<0.05)3.2The expression of Cx43, PI3K and Akt mRNA in different age andgenders3.2.1The expression of Cx43, PI3K and Akt mRNA in group of male andfemale and group of＞56and≤56years is not significant.3.3The expression of Cx43, PI3K and Akt mRNA in differentclinicopathologic features of carcinomatous gastric tissue.3.3.1The expression of Cx43mRNA in gastric carcinoma which werewell differentiated and moderately differentiated were higher than they were inpoorly differentiated gastric carcinoma (P<0.05).The expression of PI3K andAkt mRNA in gastric carcinoma which were well differentiated andmoderately differentiated was lower than that in poorly differentiated gastriccarcinoma (P<0.05).3.3.2The expression of Cx43mRNA in gastric carcinoma without lymphnode metastasis were higher than they were in gastric carcinoma with lymph node metastasis (P<0.05). The expression of PI3K and Akt mRNA in gastriccarcinoma without lymph node metastasis was lower than that in gastriccarcinoma with lymph node metastasis (P<0.05).3.3.3The expression of Cx43in gastric carcinoma with TNM Ⅰ/Ⅱ werehigher than they were in gastric carcinoma with TNM Ⅲ/Ⅳ(P<0.05). Theexpression of PI3K and Akt mRNA in gastric carcinoma with TNM Ⅰ/Ⅱ waslower than that in gastric carcinoma with TNM Ⅲ/Ⅳ (P<0.05).3.3.4The expression of Cx43in invasion of gastric carcinoma not gettingin touch with serosa were higher than they were in gastric carcinoma withgetting in touch with serosa (P<0.05). The expression of PI3K and Akt mRNAin invasion of gastric carcinoma not getting in touch with serosa was lowerthan that in gastric carcinoma with getting in touch with serosa (P<0.05).Conclusions1. Immunohistochemical positive signals of Cx43, PI3K and Akt weresignificant differences between normal gastric tissue and gastric carcinoma,also were concerned with differentiation degree, lymph node metastasis, TNMby stages and depth of invasion of gastric carcinoma, while not significantwith age and gender.2. western blot technology protein expressions of Cx43, PI3K and Aktwere significant differences between normal gastric tissue and gastriccarcinoma, also were concerned with differentiation degree, lymph nodemetastasis, TNM by stages and depth of invasion of gastric carcinoma, whilenot significant with age and gender.3. RT-PCR detection Cx43, PI3K and Akt mRNA were significantdifferences between normal gastric tissue and gastric carcinoma, also wereconcerned with differentiation degree, lymph node metastasis, TNM by stagesand depth of invasion of gastric carcinoma, while not significant with age andgender.Part Ⅱ The relationship among Cx43, PI3K and Akt in the gastriccarcinomaObjective To research the relationship among Cx43, PI3K and Akt in carcinomathrough analyzing experimental data of themMethodsTo analyse experimental data of Cx43, PI3K and Akt by the statisticalanalysis software.Results1. Immunohistochemical The correlation coefficient of Cx43and PI3Kprotein is r=-0.347(P<0.01); The correlation coefficient of Cx43and Aktprotein is r=-0.394(P<0.01); The correlation coefficient of PI3K and Aktprotein is r=0.532(P<0.01).2. Western blotting The correlation coefficient of Cx43and PI3Kprotein is r=-0.462(P<0.01); The correlation coefficient of Cx43andAktprotein is r=-0.431(P<0.01); The correlation coefficient of PI3K and Aktprotein is r=0.587(P<0.01).3. The correlation coefficient of Cx43mRNA and PI3K mRNA isr=-0.350(P<0.01); The correlation coefficient of Cx43mRNA and Akt mRNAis r=-0.297(P<0.01); The correlation coefficient of PI3K mRNA and AktmRNA is r=0.634(P<0.01).ConclusionsThe correlation of protein expression of Cx43and PI3K was negative;The correlation of expression of Cx43mRNA and PI3K mRNA wasnegative;The correlation of protein expression of Cx43and Akt was negative;The correlation of expression of Cx43mRNA andAkt mRNA wasnegative;The correlation of protein expression of PI3K and Akt was positive;The correlation of expression of PI3K mRNA andAkt mRNA was positive.