| Nowadays,gastric cancer is one of the common cancer,and the mortality is also increasing.Adenocarcinoma is the most common pathological type of gastric cancers.Its occurrence and development involve many complex mechanisms,including abnormality cell death and cell proliferation.Cell death include many kinds of forms.Apoptosis is mainly the focus of current research which is a highly regulated active cell death process and a specific cysteine aspartate protease(Caspases family)activated form of cell death.Apoptosis should be strictly controlled.Therefore,the activity of Caspase family is also strictly regulated.Caspase-8 is the promoter of death receptor which mediated apoptosis.It is abnormally expressed in many malignant tumors.TRAF2 is an important biological inhibitor of apoptosis,but the mechanism of inhibiting apoptosis is not very clear.Currently,it is found that it mainly induces the upregulation of downstream inhibitor of apoptosis by activating NF-KB pathway,thereby regulating cell apoptosis.Bid is a member of the BH3 family which promoting apoptosis Bcl-2 family.It is an important protein linking the cell death receptor pathway and mitochondrial pathway,which can expand the effect range of apoptosis mediated by death receptor pathway.There are also abnormal expression of TRAF2 and Bid in different malignant tumors.At present,there are few reports about the expression of TRAF2 in gastric adenocarcinoma tissues.At the same time,three methods are used to study the expression of TRAF2,Caspase-8 and Bid in gastric adenocarcinoma and their correlation have not been reported.Therefore,this study explored TRAF2,Caspase-8 and Bid expression in gastric carcinoma and normal gastric mucosa tissues by mmunohistochemistry SP,Western blot methods and real-time quantitative PCR.We also analyzed their relationship with the clinicopathological features in gastric adenocarcinoma,the correlation between the TRAF2,Caspase-8 and Bid expression in gastric adenocarcinoma tissues are also analyzed.They may provide molecular markers combined with early diagnosis and prognosis for patients with gastric cancer and may even provide a new target for clinical treatment.ObjectiveThe study is in order to explore the TRAF2,Caspase-8 and Bid expression in gastric carcinoma tissues and normal gastric mucosa tissues adjacent to carcinoma,and discusses the relationship between the three factors in gastric carcinoma and the clinical pathological features.As well as we analysied the correlation among the three factors.Methods1 Immunohistochemical method was used to detect TRAF2 protein,Caspase-8 protein and Bid protein expression in different gastric tissue,We explored its expression levels in gastric adenocarcinoma tissue and the relationship between the clinical pathological features,We also analyzed the correlation among TRAF2?Caspase-8 and Bid in gastric adenocarcinoma tissues.2 Western blot technique was used to detect TRAF2 protein,Caspase-8protein and Bid protein expression in different gastric tissue,We explored its expression levels in gastric adenocarcinoma tissue and the relationship between the clinical pathological features.We also analyzed the correlation among TRAF2?Caspase-8 and Bid in gastric adenocarcinoma tissues.3 qRT-PCR technique was used to detect TRAF2 mRNA,Caspase-8mRNA and Bid mRNA expression in different gastric tissue,We explored its expression levels in gastric adenocarcinoma tissue and the relationship between the clinical pathological features.We also analyzed the correlation among TRAF2?Caspase-8 and Bid in gastric adenocarcinoma tissues.Results1 Immunohistochemical results showed that the expression rates of TRAF2,caspase-8 and Bid in gastric adenocarcinoma were 76.7%,66.7%and 70%respectively.The positive expression rates of TRAF2,caspase-8 and Bid were 32.1%,90.9%and 87.3%respectively in the normal gastric mucosal tissues.Compared with adjacent to carcinoma tissues,The positive expression rate and positive expression intensity of TRAF2 protein in gastric adenocarcinoma increased,however,Caspase-8 and the Bid protein positive expression rate and positive expression intensity are lower than that in adjacent to carcinoma tissues,the difference was statistically significant(P<0.05,P<0.05,P<0.05).1.1 The expression positive rate of TRAF2 protein,caspase-8 protein and Bid protein in gastric adenocarcinoma was not related to the patients gender and age,which were not statistically significant(P>0.05,P>0.05).While there was related to the differentiation,TNM stage,lymph node metastasis and infiltration depth,and the difference was statistically significant(P<0.05,P<0.05,P<0.05,P<0.05).1.2 Analyze the relationship among of TRAF2 protein,Caspase-8protein and Bid protein expression in gastric adenocarcinoma tissuesThere was a negative correlation between the expression of TRAF2protein and Caspase-8 protein in gastric adenocarcinoma tissues,the difference was statisticaly significant(r_s=-0.630,P<0.05);In gastric adenocarcinoma,the expression of caspase-8 protein and Bid protein was positively correlated,and the difference was statistically significant(r_s=0.465,P<0.05).There was no correlation between the expression of TRAF2 protein and Bid protein in gastric adenocarcinoma,and the difference was not statistically significant(P>0.05).2 Western blot test results show that compared with normal gastric mucosa tissues,The relative expression of TRAF2 protein in gastric cancer tissues significantly increased;However,The relative expression of Caspase-8protein and Bid protein were dropped significantly,the difference had statistical significance(P<0.05,P<0.05,P<0.05).2.1 The relative expression of TRAF2 protein,caspase-8 protein and Bid protein in gastric adenocarcinoma was not related to the patients gender and age,and the difference was not statistically significant(P>0.05,P>0.05).The relative expression of the three proteins was related to the differentiation,TNM stage,lymph node metastasis and infiltration depth of gastric adenocarcinoma,and the difference was statistically significant(P<0.05,P<0.05,P<0.05,P<0.05).2.2 Analyze the relationship among of TRAF2 protein,Caspase-8protein and Bid protein expression in gastric adenocarcinoma tissuesThere was a negative correlation between the expression of TRAF2protein and Caspase-8 protein in gastric adenocarcinoma tissues,the difference was statisticaly significant(r_s=-0.682,P<0.05).In gastric adenocarcinoma,the expression of caspase-8 protein and Bid protein was positively correlated,and the difference was statistically significant(r_s=0.915,P<0.05).There was no correlation between the expression of TRAF2protein and Bid protein in gastric adenocarcinoma,and the difference was not statistically significant(P>0.05).3 qRT-PCR experiment technology test results show that compared with normal gastric mucosa tissues,TRAF2 mRNA expression is significantly ascendant in gastric adenocarcinoma tissues.However,Caspase-8 mRNA and Bid mRNA are relatively lower than that in adjacent to carcinoma tissues,the difference was statistically significant(P<0.05,P<0.05,P<0.05).3.1 The expression levels of TRAF2 mRNA,caspase-8 mRNA and Bid mRNA in gastric adenocarcinoma were not correlated with the patients gender and age,and the differences were not statistically significant(P>0.05,P>0.05).But their mRNA expression were related to the differentiation,TNM stage,lymph node metastasis and infiltration depth in gastric adenocarcinoma,and the difference was statistically significant(P<0.05,P<0.05,P<0.05,P<0.05).3.2 Analyze the relationship among of TRAF2 mRNA,Caspase-8mRNA and Bid mRNA expression in gastric adenocarcinoma tissuesThere was no obvious correlation among the TRAF2?Caspase-8 and Bid mRNA,there was no statistically significant difference(P>0.05,P>0.05,P>0.05).Conclusion1 The expression level of TRAF2 molecules in gastric adenocarcinoma tissue was higher than in normal gastric mucosa tissues at protein and mRNA level.However,Caspase-8 and Bid molecular expression were the opposite of the trend.Which all were related to the differentiation,TNM clinial stage,lymphnodemetastasisandinfiltrationdepthofgastric adenocarcinoma.TRAF2,Caspase-8 and Bid are participate in the occurrence and development of gastric adenocarcinoma,which played an important role in the evolution of the gastric adenocarcinoma,concluded the research.2 Through the analysis of the immunohistochemical SP and Western blot experimental results,the conclusion is drawn.There was a negative correlation between the TRAF2 and Caspase-8 protein,while a positive correlation between Caspase-8 protien and Bid protein.There was no statistical correlation between TRAF2 protein and Bid protein.It is suggested that there may be antagonism between TRAF2 protein and Caspase-8protien,while synergistic effects between Caspase-8 protein and Bid protein in regulating the apoptosis of gastric adenocarcinoma cells.There may be other complex apoptosis regulating signals between TRAF2 protein and Bid protein.3 Through the analysis of the qRT-PCR experimental results,the conclusion is drawn.There was no correlation among TRAF2,caspase-8 and Bid mRNA in gastric adenocarcinoma,which suggesting that there may be no direct mutual regulation between the three genes at the gene level.There may be more other regulatory pathways between them. |
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