| Objective:Screen and detect the expression level of HOXB2 in gastric adenocarcinoma,and combine with the corresponding clinicopathological data to analyze the effect of HOXB2 on the clinicopathological characteristics and survival of patients with gastric adenocarcinoma,and explore the effect of HOXB2 on the proliferation of gastric adenocarcinoma cell lines.Methods:The microarray analysis was used to screen the HOXB2 gene abnormally expression in gastric adenocarcinoma.Collected a total of 60 pairs of surgical specimens of gastric adenocarcinoma and corresponding normal gastric tissues with clear pathological diagnosis,which came from the department of gastrointestinal surgery,from January 2019 to April 2019 at Yantai Yuhuangding Hospital Affiliated to Qingdao University.Real-time quantitative PCR(RT-q PCR)and Western blot technology were used to detect the HOXB2 m RNA and protein levels in samples.Gene knockdown technology was used to establish a gastric adenocarcinoma cell line(AGS cell line)with HOXB2 knockdown,and the proliferation of AGS cell lines was observed.A total of 114 cases of surgically removed,pathologically diagnosed gastric adenocarcinoma and corresponding normal gastric tissue postoperative pathological sections were collected from January 2013 to December 2014 in Yantai Yuhuangding Hospital Affiliated to Qingdao University.Immunohistochemistry(IHC)was used to detect the expression of HOXB2 in the 114 pathological sections,and the correlation between the results with clinical pathology data and survival time was analyzed.Kaplan-Meier method was used for survival analysis,and Log-rank method was used to test.Results:1.c DNA microarray screening results show: Compared with normal gastric tissue,the expression of HOXB2 gene in gastric adenocarcinoma is 6.29 times that of normal gastric tissue(P<0.01).2.RT-q PCR results showed that the expression of HOXB2 m RNA in gastric adenocarcinoma tissues(1.329±0.484)was higher than that in normal tissues(1.129±0.582),and the difference between each other was statistically significant(P<0.05).3.Western blot results showed that the expression of HOXB2 protein in gastric adenocarcinoma tissue was(0.675±0.178)higher than that in normal tissues(0.614±0.193),and the difference was statistically significant(P<0.01).4.Analysis of immunohistochemistry results showed that the expression rate of HOXB2 in gastric adenocarcinoma tissue was 64.9%(74/114)higher than that in normal gastric tissue,28.9%(33/114),and the difference between the two was statistically significant(P<0.001);the positive rate of well differentiated adenocarcinoma in gastric adenocarcinoma group was 31.6%(6/19),the positive rate of moderately differentiated adenocarcinoma was 65.6%(40/61),and the positive rate of poorly differentiated adenocarcinoma was 82.4%(28/34),the difference between the three was statistically significant(P=0.001),suggesting that the worse the tissue differentiation,the higher the expression of HOXB2 protein.5.Univariate analysis showed that the expression level of HOXB2 protein was correlated with the depth of invasion,TNM stage,tumor size and tumor differentiation(P<0.05),and there is no correlation with age,gender,lymph node metastasis,tumor location,vascular tumor thrombus,nerve infiltration,and distant metastasis.6.Binary Logistics regression analysis showed that TNM staging and tumor differentiation were independent influencing factors of HOXB2 expression;KaplanMeier survival analysis showed that the 5-year survival rate of HOXB2 gene-positive group was 23.4%,which was lower than HOXB2 gene-negative group 42.4%(P<0.05).7.Established a cell line with HOXB2 gene knockdown;CCK8 results showed that the proliferation of the HOXB2 gene knockdown cell group was significantly slower than that of the blank control group at the 48 h,72h and 96 h of culture,and the difference was statistically significant(all P<0.05).Conclusion:The expression of HOXB2 gene in gastric cancer tissues is higher than that in normal tissues.The expression level of HOXB2 is related to TNM stage and tumor differentiation.The survival time of HOXB2 positive group is shorter,and HOXB2 gene knockdown can reduce the proliferation of gastric cancer cells.This suggests that HOXB2 may be involved in the occurrence and development of gastric adenocarcinoma,and may become a reference indicator for evaluating the prognosis of gastric adenocarcinoma. |
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