| Objective: In order to explore the expression and significance of CUL4 B and P16 in gastric carcinoma.Methods: 25 cases of gastric cancer patients who visited the second hospital of Hebei Medical University from January 2016 to December 2016 and accepted operation treatment were randomly involved in our experiment with the permission of themselves.The carcinoma tissues of the 25 cases were regarded as the experimental group,using the corresponding respected edge tissues from normal gastric tissue as the experimental control group.The application of HE staining was used to confirm gastric cancer type and determine the degree of differentiation;Immunohistochemical method and Western blot were used to detect the expression of CUL4 B protein and P16 protein in gastric cancer tissues and their corresponding normal tissues in order to explore differential expression in the experimental group and and analyze the relationship between tumor progression factors.Results:1 Case data HE staining,the histological types of gastric carcinoma tissues and their corresponding normal tissues were determined under microscope.,the type of gastric cancer: 19 cases of adenocarcinoma,2 cases of mucinous adenocarcinoma,3 cases of signet-ring cell carcinoma,1 cases of neuroendocrine carcinoma;differentiation: 3 cases of high differentiation,9cases of moderately differentiated,13 cases of poorly differentiated;more clinical data such as gender,age,clinical stage and clinical pathological factors(Table1).2 Immunohistochemical analysis of gastric tissue2.1.1 The expression of CUL4 B protein in gastric cancer tissues and theircorresponding normal tissues: according to the results of immunohistochemist-ry,it was shown that CUL4 B mainly localized in the nucleus,and the nucleus appearing clear yellow,yellow brown to dark brown color was considered as positive expression standard.CUL4 B protein in gastric tissue was positive or strong positive expression,the integral optical density(OD)of CUL4 B is0.180±0.019,whereas positive and weakly positive expression was detected in their normal counterpart with less OD value,0.128±0.012.The Dyeing intensity of CUL4 B in gastric cancer tissues was significantly higher than that in control their corresponding normal tissues,and the difference between two the groups was statistically significant(P<0.01);2.1.2 CUL4 B protein expression level in different stages of gastric cancer:CUL4B protein was positive in almost all the tissues of gastric cancer.The protein expression levels are as follows: the OD of stage I group is0.160±0.011,the OD of stage II group is 0.176±0.015,and the average OD of stage III+ IV group is 0.191±0.016.P value was 0.200 based on the t-test with the OD values of the stage I group and II group,P value was 0.021 based on the t-test with the OD values of the stage I group and stage III + IV group,P value was 0.260 based on the t-test with the OD values of the stage II group and stage III + IV group,so the discrepancy between stage I group and stage III+IV group was statistically significant(P<0.05);2.1.3 The expression of CUL4 B protein and gastric cancer’s clinical pathological factors: carcinoma tissue differentiation degree: the OD of high+medium differentiation group is 0.162±0.009,and the OD of low differentiation group is 0.192±0.013,so high+medium differentiation group of CUL4 B protein positive expression was lower than low differentiation group,and the difference between two the groups was statistically significant(P<0.05);carcinoma with invasion depth: the OD of T staging T1+T2 group is0.164±0.012,and the OD of T staging T3+T4 group is 0.191±0.016,hence CUL4 B protein expression level of T1+T2 group was significantly lower than T3+T4 group with statistically significant difference(P<0.01);The statistical analysis of other factors such as sex,age,the histological types,lymph nodemetastasis and remote metastasis fail to indicate significant difference.2.2.1 The expression of P16 protein in gastric cancer tissues and their corresponding normal tissues: The major positive location of P16 protein staining was in the nucleus.The expresssion level of P16 protein in gastric tissue varies from negative,weakly positive to positive expression,the over all OD value of P16 in gastric cancer group is 0.112±0.017,whereas positive and strong positive expression in their normal counterparts were detected with OD value of 0.151 ±0.012.The Dyeing intensity of P16 in gastric cancer group was significantly lower than that in control group with statistically significant difference(P<0.01);2.2.2 P16 protein expression level in different stages of gastric cancer: P16 protein was mostly negative or weakly positive in gastric cancer.The protein expression levels are as follows: the OD of stage I group is 0.129±0.009,the OD of stage II group is 0.120±0.006,and the average OD of stage III+IV group is 0.100±0.015.P value was 0.229 based on the t-test with the OD values of the stage I group and II group,P value was 0.018 based on the t-test with the OD values of the stage I group and stage III+IV group,P value was0.026 based on the t-test with the OD values of the stage II group and stage III+IV group,so the discrepancy between stage I group and stage III+IV group and the discrepancy between the stage II group and stage III+IV group were statistically significant(P<0.05);2.2.3 The expression of P16 protein and gastric cancer’s clinical pathological factors: carcinoma tissue differentiation degree: the OD of high+medium differentiation group is 0.126±0.011,and the OD of low differentiation group is 0.102±0.016,the high+ medium differentiation group of P16 protein positive expression was higher than low differentiation group,and the difference between two the groups was statistically significant(P<0.05);carcinoma with invasion depth: the OD of T staging T1+T2 group is0.126±0.009,and the OD of T staging T3+T4 group is 0.100±0.015,so T staging T1+T2 group of P16 protein positive expression was significantly higher than T3+T4 with statistically significant difference(P<0.01);Thestatistical analysis of other factors such as sex,age,the histological types,lymph node metastasis and remote metastasis fail to indicate significant difference.2.3 Correlation between the expression of CUL4 B and P16 in gastric carcinoma: Linear regression method was used to analyze the correlation between the expression of CUL4 B protein and P16 protein in gastric carcinoma by immunohistochemistry,the expression of CUL4 B and P16 in gastric carcinoma was negative correlation(r=-0.552,P<0.05).3 Western blot results of gastric tissue3.1.1 The expression of CUL4 B protein was detected by Western blot:The expressions of CUL4 B protein were(0.734±0.150)in gastric cancer tissues,(0.374±0.052)in their corresponding normal tissues.The results showed that the expression of CUL4 B protein in gastric cancer tissues were higher than that in their corresponding normal tissues.There is significance statistical difference between the two groups(P<0.01);3.1.2 The expression of CUL4 B protein and gastric cancer’s clinical staging:The quantized expression of CUL4 B protein was 0.577±0.035 in stage I group,0.642±0.025 in stage II group,0.860±0.099 in III+IV group.P value was0.061 between the stage I group and II group,P value was 0.000 between the stage I group and stage III+IV group,P value was 0.002 between the stage II group and stage III+IV group,so the difference between stage I group and stage III+IV group,and the difference between stage II group and stage III+IV group were statistically significant(P<0.01);3.1.3 The expression of CUL4 B protein and gastric cancer’s clinical pathologi-cal factors: carcinoma tissue differentiation degree: The expression of CUL4 b protein were 0.614±0.070 in high+ medium differentiation group,0.820±0.131 in low differentiation group,the results show that the expression of CUL4 B protein in high+ medium differentiation group was lower than that in low differentiation group,and the difference between two the groups was statistically significant(P<0.05);carcinoma with invasion depth: The expression of CUL4 B protein was 0.609±0.045 in T staging T1+T2 group,0.860±0.099 in T3+T4 group,so the expression of CUL4 B protein in T1+T2group was lower than that in T3+T4 group,and the difference between two the groups was statistically significant(P<0.01);The statistical analysis of other factors such as sex,age,the histological types,lymph node metastasis and remote metastasis fail to indicate significant difference.3.2.1 The expression of P16 protein was detected by Western blot:The expression of P16 protein was 0.553±0.096 in gastric cancer tissues,0.894±0.187 in their corresponding normal tissues.The results showed that the expression of P16 protein in gastric cancer tissues was lower than that in their corresponding normal tissues.There is significance statistical difference between the two groups(P<0.01);3.2.2 The expression of P16 protein and gastric cancer’s clinical staging:The expression of P16 protein was 0.643±0.042 in stage I group,0.601±0.069 in stage II group,0.484±0.075 in stage III+IV group.P value was 0.411 between the stage I group and II group,P value was 0.012 between the stage I group and stage III+IV group,P value was 0.059 between the stage II group and stage III+IV group,so the difference between stage I group and stage III+IV group were statistically significant(P<0.05);3.2.3 The expression of P16 protein and gastric cancer’s clinical pathological factors: carcinoma tissue differentiation degree: The expression of P16 protein was 0.629±0.054 in high+medium differentiation group,with 0.498±0.082 in low differentiation group,so the results showed that the expression of P16 protein in high+medium differentiation group was higher than that in low differentiation group,and the difference between two the groups was statistically significant(P<0.05);carcinoma with invasion depth: The expressi-on of P16 protein was 0.621±0.056 in T staging T1+T2 group,and0.484±0.075 in T3+T4 group,so the expression of P16 protein in T1+T2group were higher than that in T3+T4 group,and the difference between two the groups was statistically significant(P<0.01);The statistical analysis of other factors such as sex,age,the histological types,lymph node metastasisand remote metastasis fail to indicate significant difference.3.3 The correlation analysis of the expression of CUL4 B and P16 in gastric cancer tissues:the expression of CUL4 B and P16 in gastric cancer group was negative correlation(r=-0.667,P<0.05).Conclusion:1 The positive expression of CUL4 B protein in gastric cancer tissues was evidently higher than their corresponding normal tissues,Positive correlation of CUL4 B protein expression with gastric cancer clinical stage,depth of invasion and differentiation revealed its involvement in the process of malignant transformation of gastric cancer.Elevated expression of CUL4 B in gastric cancer cells indicates its potential utility as one of the gastric cancer tumor marker,and our test provided the experimental basis for the evaluation and clinical diagnosis of gastric cancer.2 The expression of P16 protein in gastric cancer was decreased,which was negatively correlated with the clinical stage,depth of invasion and differentiation of gastric cancer.It was shown that P16 could inhibit the occurrence and development of gastric cancer as a tumor suppressor gene.3 The expression of CUL4 B and P16 were negatively correlated. |
