Study Of MiR-410 Targeting Snai1 And Regulates Emt And Inhibits Invasion Of Gastric Carcinoma Cells

Objective: To investigate the relationship between mi R-410 and the epithelial mesenchymal trasition(EMT),and analyze the relationship between EMT key factor Snai1 and mi R-410.And clarify the relationship between mi R-410 targeting Snai1 and regulates EMT and the invasion in gastric carcinoma.Methods: Firstly,12 mi RNAs were found to combin with Snai1 through mi Rwalk online progress and mi R-410 was found to be the best binding mi RNA of Snai1.The expression level of Snai1 was detected by immunohistochemistry in 75 cases of gastric carcinoma tissues and 30 cases of Gastric mucosa adjacent to carcinoma.The relationship between Snai1 and Clinicopathological features of gastric carcinoma were analysized.And also the expressions of mi R-410 and Snai1 were detected with q RT-PCR and Western blot,respectively,and the correlation between mi R-410 and Snai1 were analysized.Expression of mi R-410 was detected by q RT-PCR and expressions of Snai1,E-cadherin and Vimentin were detected by Western blot in different differentiation degree gastric carcinoma cell lines and the relationship among mi R-410 and three proteins were analysized.In the recombinant cell lines?SGC-7901-mi R-410 mimics ? SGC-7901-mi R-NC ?SGC-7901-mi R-410-Inhibitors?SGC-7901,q RT-PCR were used to detect the expressions of mi R-410 and western blot were used to detect the expressions of Snai1,E-cadherin and Vimentin,respectively.And we analysized whether mi R-410 could bind to the 3` UTR site of Snai1.The changes of invasive ability of recombinant gastric cancer cell lines after changed expression of miR-410 were detected by transwell method.Results:(1)12 mi RNAs were found to combine with Snai1 through bioinformatics methods and among these im RNAs,mi R-199a-5p and mi R-410 had the the best specificity and the strongest stability for combine with Snai1.(2)Positive expression rate of Snai1 was 82.7%(62/75)in gastreic carcinoma tissues and that 23.3%(7/30)in Gastric mucosa adjacent to carcinoma.There were significant difference between them(P<0.01).(3)mi R-410 had lower expression in gastric carcinoma tissues than in Gastric mucosa adjacent to carcinoma,while Snai1 had higher expression in gastric carcinoma tissues than in Gastric mucosa adjacent to carcinoma(P<0.01).There were inverse relation between mi R-410 and Snai1.(4)Positive expression rate of Snai1 in low differentiated gastric carcinoma was 93.2%(41/44)and it was higher than 67.7%(21/31)in high differentiated gastric carcinoma.Positive expression rate of Snai1 in clinical stage III+IV was 92.9%(39/42)and it was higher than 69.7%(23/33)in clinical stage I+II.Positive expression rate of Snai1 in patients with lymph nodes metastasis was 93.6%(44/47)and it was higher than 64.3%(18/28)in patients without lymph node metastasis.Positive expression rate of Snai1 in patients with distant metastasis was 92.5%(37/40)and it was higher than 71.4%(25/35)in patints without distant metastasis.And the above differences were statistically significant(P < 0.05)(5)In low differentiated gastric carcinoma cell line,there were low expression of mi R-410 and high expression of Snai1.In middle and well differentiated gastric carcinoma cell line,there were hihg expression of mi R-410 and low expression of Snai1.There maybe relationship among high expression of mi R-410 and low expression of Snai1 and the occurrence and development of gastric carcinoma.(6)mi R-410 was increased in cell line SGC-7901-mi R-410 mimics and Snai1 was downregulated and lead to E-cadherin upregulation and Vimentin downregulation.mi R-410 was decreased in cell line SGC-7901-mi R-410 inhibitors and Snai1 was upregulated and lead to E-cadherin downregulation and Vimentin upregulation.(7)The count of invasive cells in SGC-7901-mi R-410 mimics were more than other three groups cell line.The count of invasive cells in SGC-7901-mi R-410 inhibitors were less than other three groups cell line (P<0.01).There were no significant difference between the count of invasive cells of SGC-7901-mi R-NC cell line and SGC-7901 cell line(P>0.05).Conclusions:(1)There were high expression of Snai1 and low expression of mi R-410 in gastric carcinoma tissues,and that relating to the occurrence and development of gastric carcinoma.(2)mi R-410 can target Snai1 and downregulate expression of Snai1 and regulates the EMT signaling pathway and inhibits invasion ability of gastric carcinoma cells.