| Objective: This study was to investigate the expression of the miRNA-320 d and FoxM1 in the gastric cardia adenocarcinoma(GCA),and to explore their association with clinical pathological characteristics and prognosis of cardia cancers.In addition,we through this study to detect the influence of mi RNA-320 d and Fox M1 in the process of the development of GCA,to analyse the potential regulation and control relations between the miRNA-320 d and Fox M1,to explore the mechanism of their effect on occurrence and development of GCA.Methods: We collected 30 cases of cardia cancer patients admitted in the First Affiliated Hospital of Henan University of Science and Technology in the period from April 2012 to March 2013,and 30 cases of patients with cardiac cancer admitted in Anyang Tumor Hospital from March 2013 to August 2013.We collected cardiac carcinoma tissues and matching tissues adjacent to carcinoma when the radical resection of the cardia cancer was performed,all cases were without preoperatively radiotherapy and chemotherapy.Gene primers of miRNA-320d/FoxM1 were designed and RT-PCR was used to determine the level of mi RNA-320d/FoxM1 m RNA expressed in GCA and matching tissues adjacent to carcinoma respectively.The expression level of FoxM1 protein in GCA and matching tissues adjacent to carcinoma was detected by immune histochemical method and Western blot.Analysis of the relationship between mi RNA-320d/FoxM1 expression and clinicopathological features and prognosis of patients with cardia cancer.Using Graphpad Prism5.0 statistical software for statistical data analysis,and statistical results with P<0.05 for the difference was statistically significant.Results: 1.RT-PCR showed that the mRNA expressions of mi RNA-320 d in carcinoma of gastric cardia tissues were significantly lower than that in normal tissues adjacent to carcinoma(P<0.001);The expressions of Fox M1 on mRNA level in cardiac carcinoma tissues were higher than that in matching tissues adjacent to carcinoma(P<0.0001).We examined the relationship between the m RNA expressions of miRNA-320d/FoxM1 and patients clinical parameters.Analysis showed that miRNA-320 d expression was irrelevant with age,sex,tumor differentiation,lymphnode metastasis and TNM stage,but was in a positive correlation with the tumor size(P<0.05).FoxM1 expression was irrelevant with age,sex,tumor differentiation and lymphnode metastasis,but was in a positive correlation with the tumor size and TNM stage(P<0.05),with significant statistical difference.Then we analyzed the relationship between mi RNA-320 d and FoxM1 expression and found that the expression of miRNA-320 d and Fox M1 were negatively correlated in nucleic acid level.2.Western blot showed that the expressions of FoxM1 on protein level in cardiac carcinoma tissues were higher than that in matching tissues adjacent to carcinoma(P =0.0363),with significant statistical difference.3.Immunohistochemistry indicated that the positive expression of FoxM1 was brown-yellow granules,mainly located in the nucleus,part of the cytoplasm can also be seen coloring.The positive rate of FoxM1 expression was 68% in GCA and 45% in the adjacent non-cancerous tissues,with significant statistical difference(P<0.01).We examined the relationship between the positive expression of FoxM1 and patients clinical parameters.Analysis showed that the positive expression of FoxM1 was in a positive correlation with the tumor size and TNM stage(P<0.05).Then we studied the expression of RT-PCR?WB and IHC,the outcomes indicated that FoxM1 mRNA and protein expression trends were accordance.4.Then our study group followed up the 60 patients with cardia cancer.KaplanMeier survival analysis showed that the overall survival(OS)of cardiac carcinoma patients who have high expression of miRNA-320 d is higher than that of low expression(P<0.01);The overall survival(OS)of cardiac carcinoma patients who have low expression of FoxM1 is higher than that of high expression(P <0.01).Conclusions: 1.The expression of mi RNA-320 d in GCA was significantly lower than that in the adjacent non-cancerous tissues,and the expression of Fox M1 in GCA was significantly higher than that in the adjacent non-cancerous tissues,suggesting that the two may be involved in the occurrence and development of GCA,and FoxM1 may be the potential downstream target gene of miRNA-320 d in carcinoma of gastric cardia.However,the specific mechanism remains to be further explored.2.Through correlating with clinical pathological features,the mi RNA-320 d expression was related to tumor size,and expression of Fox M1 was in a positive correlation with the tumor size and TNM stage.The OS of cardiac carcinoma patients who have high expression of miRNA-320 d is higher than that of low expression;FoxM1 positive expression group had a poorer prognosis compared with negative expression group.So both of these is expected as molecular targets in the prognosis of cardiac carcinoma patients. |
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