A Preliminary Study Of The Expression And Regulation Of MiR-449a/b And Notch1in NSCLC Tissues

Objective：Investigating the expression of Notch1and miR-449a/b in NSCLC tissues andanalysis the correlation,to explore miR449a/b targeted effects on notch1.Methods：miR-449a was evaluated by quantitative real time qRT-PCR,and the expression ofNotch1mRNA and protein were assessed by RT-PCR and westernblotting,respectively.Finally,validating the targeting relationship between miR-449a/band notch1’s by Biological Analysis and luciferase assay.①In90cases tissue samples, carcinoma notch1positive expression rate was73.3%; tissue beside carcinoma was30%, the difference was statistically significant.（P＜0.05）。②In this experiment, Notch1protein expression and clinicopathologicalfactors correlation analysis found that there are differences in different pathologicaltypes of expression of Notch-1, the difference was statistically significant (P<0.05).Further, in conjunction with other clinical and pathological analysis notch1 various factors, it was found significantly associated with TNM stages expression ofNotch-1(P <0.05)However, analysis of the age factor, gender, tumor location, sizeand lymph node metastasis showed no correlation between areas (P>0.05).③Samples with high expression of miR-449a has42cases, including21casesof carcinoma,21cases in adjacent tissue, statistical analysis of their expression incancer tissues and adjacent tissues without significant difference.And Samples withhigh expression of miR-449b,39cases, including18cases of carcinoma,21cases ofadjacent tissues. Statistical analysis of their expression in cancer tissues and adjacenttissues without significant difference.These results suggest that high expression ofmiR-449a/b have no significant correlation with lung cancer.In addition, theexpression of miR-449a/b compared with the results of Notch1protein expressionmatching is poor.④The Dual luciferase reporter gene assay results indicate that the regulatoryrole of miR-449a/b on Notch13’UTR is unobvious,there was no significantdifference.Conclusions：①Notch1protein compared to adjacent tissue is highly expressed in lung cancertissues, suggesting that in lung cancer, notch1expression levels higher than theadjacent tissues and may be associated with tumor development.notch1mRNAexpression levels of the test results also support this conclusion;②In lung squamous cell carcinoma notch1protein expression was significantlyhigher than in lung adenocarcinoma and small cell lung cancer, the difference wasstatistically significant,suggesting that notch1histological types of lung cancer maybe related to the patient may be closely related to lung cancer cells.③Lower expression notch1in I, II than III, IV in lung cancer patients,whichTips notch1may be related to cancer invasion;④MiR-449a/b expression levels in lung tissue showed no correlation with lungcancer;⑤MiR-449a is poor in comparison with the matching Notch1, showed no significant correlation. The futher dual luciferase reporter gene assay results indictthat the regulatory role of miR-449a/b on Notch13’UTR is unobvious.