| Gastric carcinoma is one of the malignant tumors of the digestive tract,and most of it is gastric adenocarcinoma,which poses a serious threaten to human health,and it is not fully clear about its causes and mechanisms.It has been fund that the abnormal mechanism of apoptosis plays a more and more important role in the development and progression of malignant tumors.Apoptosis,also known as “type I programmed cell death”,is an important form of programmed cell death,which is tightly regulated by a variety of genes.Caspase-3 and Caspase-9 are the key molecules involved in apoptosis in the Caspases family,which is an essential component of apoptosis mechanism,and c-IAP1 is an important molecule endogenous inhibitors of apoptosis in the IAPs family which is involved in apoptosis inhibition by various ways,they have abnormal expression in many human malignant tumors.The study of c-IAP1 in gastric adenocarcinoma tissue has not been reported in China,and the correlation with caspase-3 and Caspase-9expression has not been reported.The study of Caspase-3 and Caspase-9 in gastric adenocarcinoma which is limited to protein level,at the same time and the correlation between Caspase-3 and Caspase-9 have not been reported.The expression of Caspase-3 and Caspase-9 in malignant tumors such as gastric adenocarcinoma is controversial.Therefore,we examined the protein and m RNA expression of Caspase-3,Caspase-9 and c-IAP1 in gastric adenocarcinoma tissues and normal gastric mucosa tissues to explore their role in the development and progression of gastric adenocarcinoma.We analyzed their relationship with the clinicopathological features of gastric adenocarcinoma and the correlation between the Caspase-3 protein,Caspase-9protein and c-IAP1 protein expression in gastric adenocarcinoma tissues,so asto provide new molecular targets for early diagnosis,prognosis and clinical targeted therapy for gastric adenocarcinoma patients.ObjectiveThis study was designed to investigate the protein and m RNA expression of apoptosis-related gene Caspase-3,Caspase-9 and c-IAP1 in gastric adenocarcinoma tissues and normal gastric mucosa tissues to explore their role in the development and progression of gastric adenocarcinoma and relationship with the clinicopathological features of gastric adenocarcinoma.We analyzed the correlation between the Caspase-3 protein,Caspase-9 protein and c-IAP1 protein expression in gastric adenocarcinoma tissues.Methods1.A total of 50 cases of new diagnosed gastric adenocarcinoma patients who underwent surgical resection and confirmed by pathology,and 40 cases of normal gastric mucosa tissues which was range 10 cm from the edge of gastric adenocarcinoma were collected in many hospitals from October 2014 to December 2015 in Chengde city,Hebei Provience.2.Immumohistochemical method was used to detect the protein expression of Caspase-3,Caspase-9 and c-IAP1 in gastric adenocarcinoma tissues and normal gastric mucosa tissues.We analyzed the relationship between the protein expression of Caspase-3,Caspase-9 and c-IAP1 and the clinicopathological features,the correlation between the Caspase-3 protein,Caspase-9 protein and c-IAP1 protein expression in gastric adenocarcinoma tissue.3.The protein expression of Caspase-3,Caspase-9 and c-IAP1 in gastric adenocarcinoma tissues and normal gastric mucosa tissues was evaluated by Western blot.We analyzed the relationship between the protein expression of Caspase-3,Caspase-9,c-IAP1 and the clinicopathological features in gastric adenocarcinoma tissues.4.The m RNA expression of Caspase-3,Caspase-9 and c-IAP1 in gastric adenocarcinoma tissues and normal gastric mucosa tissues was examined byreal-time quantitative PCR(q RT-PCR).We analyzed the relationship between the m RNA expression of Caspase-3,Caspase-9 and c-IAP1 and the clinicopathological features in gastric adenocarcinoma tissues.5.The SPSS19.0 statistical software was used for statistical analysis,and then drawing the experimental conclusion.Results1 Immunohistochemical test results:The positive expression rates of Caspase-3 protein and Caspase-9 protein in gastric adenocarcinoma tissues were 34.0% and 30.0%,respectively,which were significantly lower than those in normal gastric mucosa tissues(85.0% and 82.5%,respectively),the difference was statistically significant(P<0.05,P<0.05).The positive expression rate of c-IAP1 in gastric adenocarcinoma tissues(76.0%)was significantly higher than that in normal gastric mucosa tissues(27.5%),the difference was statistically significant(P<0.05).1.1 The positive expression of Caspase-3 protein and Caspase-9 protein in gastric adenocarcinoma tissues were correlated with histological differentiation,TNM clinical stage and lymph node metastasis,which was significantly different(P<0.05,P<0.05,P<0.05),while there was no correlation with gender,age and depth of invasion,the difference was not statistically significant(P>0.05,P>0.05,P>0.05).The positive expression of Caspase-3 protein and Caspase-9 protein in well-moderately differentiated group,TNM clinical stage I-II group and non-lymph node metastasis group in gastric adenocarcinoma tissues was significantly higher than that in poorly differentiated group,TNM clinical stage III-IV group and lymph node metastasis group,the difference was statistically significant(P<0.05,P<0.05,P<0.05).1.2 The positive expression of c-IAP1 protein in gastric adenocarcinoma tissues was correlated with histological differentiation,TNM clinical stage,lymph node metastasis and depth of invasion,which was significantly different(P<0.05,P<0.05,P<0.05,P<0.05),while there was no correlation with gender and age,the difference was not statistically significant(P>0.05,P>0.05).The positive expression of c-IAP1 protein in well-moderately differentiated group,TNM clinical stage I-II group,non-lymph node metastasis group and non-infiltratingserosal layer in gastric adenocarcinoma tissues was significantly lower than that in poorly differentiated group,TNM clinical stage III-IV group,lymph node metastasis group and infiltrating serous layer,the difference was statistically significant(P<0.05,P<0.05,P<0.05,P<0.05).1.3 Analyze the relationship of Caspase-3 protein,Caspase-9 protein and c-IAP1 protein expression in gastric adenocarcinoma tissues1.3.1 There was a positive correlation between the expression of Caspase-3 protein and Caspase-9 protein in gastric adenocarcinoma tissues,the difference was statisticaly significant(rs=0.608,P<0.05);1.3.2 There was a negative correlation between the expression of Caspase-3 protein and c-IAP1 protein in gastric adenocarcinoma tissues,the difference was statisticaly significant(rs=-0.613,P<0.05);1.3.3 There was a negative correlation between the expression of Caspase-9 protein and c-IAP1 protein in gastric adenocarcinoma tissues,the difference was statisticaly significant(rs=-0.578,P<0.05).2 Western blot test results: The relative expression of Caspase-3 protein and Caspase-9 protein in gastric adenocarcinoma tissues were significantly lower than that in normal gastric mucosa tissues,which was significantly different(P<0.05,P<0.05).The relative expression of c-IAP1 protein in gastric adenocarcinoma tissues was significantly higher than that in normal gastric mucosa tissues,which was significantly different(P<0.05).2.1 The relative expression of Caspase-3 protein and Caspase-9 protein in gastric adenocarcinoma tissues was correlated with histological differentiation,TNM clinical stage and lymph node metastasis,which was significantly different(P<0.05,P<0.05,P<0.05),while there was no correlation with gender,age and depth of invasion,the difference was not statistically significant(P>0.05,P>0.05,P>0.05).The relative expression of Caspase-3 protein and Caspase-9 protein in well-moderately differentiatedgroup,TNM clinical stage I-II group and non-lymph node metastasis group in gastric adenocarcinoma tissues was significantly higher than that in poorly differentiated group,TNM clinical stage III-IV group and lymph node metastasis group,the difference was statistically significant(P<0.05,P<0.05,P<0.05).2.2 The relative expression of c-IAP1 protein in gastric adenocarcinoma tissues was correlated with histological differentiation,TNM clinical stage,lymph node metastasis and depth of invasion,which was significantly different(P<0.05,P<0.05,P<0.05,P<0.05),while there was no correlation with gender and age,the difference was not statistically significant(P>0.05,P>0.05).The relative expression of c-IAP1 protein in well-moderately differentiated group,TNM clinical stage I-II group,non-lymph node metastasis group and non-infiltrating serosal layer in gastric adenocarcinoma tissues was significantly lower than that in poorly differentiated group,TNM clinical stage III-IV group,lymph node metastasis group and infiltrating serous layer,the difference was statistically significant(P<0.05,P<0.05,P<0.05,P<0.05).3 Real-time quantitative PCR(q RT-PCR)test results:The m RNA expression of Caspase-3 and Caspase-9 in gastric adenocarcinoma tissues were significantly lower than that in normal gastric mucosa tissues,which was significantly different(P<0.05,P<0.05).The expression of c-IAP1 protein in gastric adenocarcinoma tissues was significantly higher than that in normal gastric mucosa tissues,which was significantly different(P<0.05).3.1 The m RNA expression of Caspase-3 and Caspase-9 in gastric adenocarcinoma tissues was correlated with histological differentiation,TNM clinical stage and lymph node metastasis,which was significantly different(P<0.05,P<0.05,P<0.05),while there was no correlation with gender,age and depth of invasion,the difference was not statistically significant(P>0.05,P>0.05,P>0.05).The m RNA expression of Caspase-3 and Caspase-9 in well-moderately differentiated group,TNM clinical stage I-II group and non-lymph node metastasis group in gastric adenocarcinoma tissueswas significantly higher than that in poorly differentiated group,TNM clinical stage III-IV group and lymph node metastasis group,the difference was statistically significant(P<0.05,P<0.05,P<0.05).3.2 The m RNA expression of c-IAP1 in gastric adenocarcinoma tissues was correlated with histological differentiation,TNM clinical stage,lymph node metastasis and depth of invasion,which was significantly different(P<0.05,P<0.05,P<0.05,P<0.05),while there was no correlation with gender and age,the difference was not statistically significant(P>0.05,P>0.05).The m RNA expression of c-IAP1 in well-moderately differentiated group,TNM clinical stage I-II group,non-lymph node metastasis group and non-infiltrating serosal layer in gastric adenocarcinoma tissues was significantly lower than that in poorly differentiated group,TNM clinical stage III-IV group,lymph node metastasis group and infiltrating serous layer,the difference was statistically significant(P<0.05,P<0.05,P<0.05,P<0.05).Conclusion1 The low expression of Caspase-3 protein and m RNA,Caspase-9protein and m RNA in gastric adenocarcinoma tissues were involved in the development and progression of gastric adenocarcinoma,which were correlated with the clinicopathological features of gastric adenocarcinoma,including histological differentiation,TNM clinical stage and lymph node metastasis,while there was no correlation with the gender,age and depth of invasion.The high expression of c-IAP1 protein and m RNA in gastric adenocarcinoma tissues was involved in the development and progression of gastric adenocarcinoma,which was correlated with the clinicopathological features of gastric adenocarcinoma,including TNM clinical stage,lymph node metastasis and depth of invasion,while there was no correlation with the gender and age.2 There was a positive correlation between the expression of Caspase-3and Caspase-9 in gastric adenocarcinoma tissues;The expression of c-IAP1 was negatively correlated with the expression of Caspase-3 protein and Caspase-9 in gastric adenocarcinoma tissues.3 Dysregulation expression of Caspase-3,Caspase-9 and c-IAP1 in gastric adenocarcinoma tissues participated in the development and progression of gastric adenocarcinoma,c-IAP1 may inhibit the apoptosis of gastric adenocarcinoma cell by down-regulated the expression of Caspase-3and Caspase-9,which contribute to the development and progression of gastric adenocarcinoma. |
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