| Background: Gastric cancer is one of the most important cancers in the worldwide which has a high mortality rate.And it is accounting for the third leading cause about the factor of cancer.The 5-yearpostoperative survival rate after gastric cancer resection is varying,Early gastric cancer can reach 90%,but the rate of patients with advanced gastric cancer is close to 10%.Therefore,early diagnosis is extremely important for patients.Larger amounts of studies have shown that circRNAs may serve as new tumor-associated biomarkers for gastric adenocarcinoma.So our task screens and validates target circRNA which associated with gastric adenocarcinoma,and makes preliminary study about its clinical significance and functions.Methods:（1）High throughput microarray technology was used to screen differentially expressed circRNAs among gastric adenocarcinoma and its matched adjacent tissues in our previous study,and circRNA molecule which had not been reported in the literature,and had more than 2 fold changes in expressed levels between cancer and precancerous tissues was selected as target circRNA;（2）Detection of the expression level of Hsacirc0000390 in tissues: we detected the expression level of target circRNA Hsacirc0000390 in 80 gastric adenocarcinoma tissues and their matched paracancerous tissues by using Real-time quantitative PCR（RT-qPCR）,and verify whether the results were consistent with the chip screening results.The clinical pathological data were analyzed statistically.The expressed levels of this circRNA were then related to clinical and pathological parameter by statistical analysis;（3）Constructing lentiviral overexpression vector of target circRNA,using lentiviral infection technology to overexpress circRNA Hsacirc0000390 in gastric adenocarcinoma cells,by this way increasing its expression level by more than two times;（4）The effects of up-regulation of circRNA Hsacirc0000390 expression on the affecting biological function of gastric adenocarcinoma cells were investigated by CCK-8 assay,plate cloning assay,and Transwell assay.Results:Among 713 circRNAs differentially expressed in gastric adenocarcinoma tissues identified by our previous microarray assay,the circRNA Hsacirc0000390 was selected as target circRNA for further investigation in this study,which exhibited low expressed in gastric adenocarcinoma tissues with a 12.44 fold-changes by chip assay.RT-q PCR results further showed that the expression of Hsacirc0000390 in gastric adenocarcinoma tissues was significantly down-regulated（the proportion is 81.3%）,compared with adjacent parcancerous tissues,which levels in gastric adenocarcinoma tissues was reduced 4.85 times.The expression of Hsacirc0000390 in gastric adenocarcinoma tissues was lower by comparison（P < 0.001）.（2）The expression level of circRNA Hsacirc0000390 in gastric adenocarcinoma tissues was correlated with the N stage of TNM stage（P < 0.05）,however,not related with sex,age,location of tumors and diameter of tumors（P > 0.05）;（3）Lentivirus transduction could up-regulate the expression of circRNA Hsacirc0000390 in gastric adenocarcinoma cell lines.Up-regulation of 000390 led to inhibit the proliferation,migration and colony formation ability of gastric adenocarcinoma cells（P < 0.05）.Conclusion: The expression level of circRNA Hsacirc0000390 in gastric adenocarcinoma tissues is down-regulated compared with its matched adjacent tissues.The expression level of Hsacirc0000390 was correlated with patients’ N stage of TNM stage in gastric adenocarcinoma tissues.Up-regulation of circRNA Hsacirc0000390 expression could inhibit the biological behaviour of gastric adenocarcinoma cells,included the proliferation,migration and colony formation.Therefore,circRNA Hsacirc0000390 can be regarded as a potential biomarker for gastric adenocarcinoma. |
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