| Part â… Expression of miR-210 in gastric carcinoma and its correlation with clinicopathological factors and prognosisObjective:To investigate the clinical significance of microRNA-210 (miR-210) expression in gastric cancer (GC).Methods:The expression of miR-210 was detected in 78 cases of gastric cancer tissues and the corresponding adjacent tissues. The relationship between the expression of miR-210 and clinical pathological factors and prognosis were analyzed.Results:Real time quantitative PCR showed that the expression of miR-210 in gastric cancer was higher than that in adjacent tissues, and there was a significant difference between the two groups (P<0.05). The expression of MiR-210 was closely related to tumor size (P=0.022), lymph node metastasis (P=0.018) and TNM stage (P=0.034). The 5 years survival rate of patients with low miR-210 expression was 48.2%, and the 5 years survival rate of patients with high miR-210 expression was 30.4%, the difference had statistical significance (2=4.216, P= 0.040).Conclusion:1.Compared with the adjacent tissues, the expression of miR-210 in gastric carcinoma was significantly increased, suggesting that miR-210 may be related to the occurrence and development of gastric cancer.2.The level of miR-210 expression was related to the clinical stage and metastatic potential of gastric cancer, and it could be used as a reliable indicator of prognosis.PartnEffect of hypoxia on invasion and metastasis of gastric cancer and the role of miR-210Objective:To investigate the effect of hypoxia on proliferation, invasion and EMT of gastric cancer cells, and to study the possible role of miR-210 in this process.Methods:We use the Lipofectamine 2000 transfected hsa-mir-210 inhibitor into gastric cancer cell lines, and knocked down mir-210 expression. Real time PCR was used to detect the mir-210 expression under normoxia,hypoxia and hypoxia and inhibition status.MTT assay, Transwell invasion assay and Western blot used to detect the changes of proliferation, invasion ability and EMT related proteins.Results:Compared with the normoxia group, the expression of miR-210 in hypoxia group was significantly higher (P<0.05), and the relative expression of miR-210 in hypoxia and inhibition group was significantly lower than that in hypoxia group (P <0.05). Cells growth inhibitory rate was detected by MTT, the invasion ability was detected by transwell assay, EMT related protein expression was detected by western blot. Compared to the normoxia group, the relative survival rate and invasion cell number were significantly increased in the hypoxia group (P< 0.05); After inhibition of mir-210, the relative cell survival rate and invasion cell number were significantly reduced (P< 0.05). Western blot results showed that compared with the normoxia group, the expression of epithelial marker decreased and the expression of mesenchymal marker increased in the hypoxia group, while inhibited the expression of miR-210, the expression of EMT related protein could be reversed.Conclusion:1. Hypoxia could induce the expression of miR-210, and enhance the proliferation and invasion of gastric cancer cells, and also promote the process of EMT. 2. Knockdown of miR-210 inhibited the proliferation and invasion of gastric cancer under hypoxia status, and inhibited the EMT process.Part IIITargets of miR-210 in gastric cancer cells with hypoxia conditionObjective:To study the targets of miR-210 in gastric cancer cells with hypoxia condition.Methods:We used bioinformatics software to predict the target of miR-210, and the possible target genes and proteins in the normoxia group, hypoxia group and hypoxia inhibition group were also detected.Results:PicTar, TargetScan and mirbase were used to predict the target gene of miR-210, combined with previous literature, we found that HOXA9, EFNA3 and FGFRL1 were possible target genes. Quantitative real-time PCR results showed that the relative expression of HOXA9 in hypoxia group was lower than that of the normoxia group, and the relative expression of HOXA9 in hypoxia inhibition group was higher than that of the hypoxia group, the difference was statistically significant (P< 0.05);relative expression of FGFRL1 in hypoxia inhibition group was significantly higher than that in the hypoxia group(P<0.05). The expression of EFNA3 had no statistically significant difference among the three groups. Compared to the normoxia group, the expression of FGFRL1 protein was decreased in the hypoxia group, the difference was statistically significant (p<0.05), when compared to the hypoxia group, the expression of FGFRL1 protein was significantly increased in the hypoxia inhibition group(P<0.05). The expression of HOXA9 and EFNA3 proteins had no significant difference among the three groups.Conclusion:1. The expression of FGFRL1 was affected by hypoxia, while inhibiting the expression of miR-210 could up-regulate the expression of FGFRL1 in human gastric cancer cells.2. FGFRL1 may be the target gene of miR-210 in gastric cancer cells with hypoxia condition, and HOXA9 and EFNA3 were not the target genes of miR-210 in gastric cancer cells. |
PDF Full Text Request