Screening And Functional Mechanisms Of Non-coding RNA In Lung Adenocarcinoma

Objectives 1 To detect the difference of expression level of circRNA hsa_circ₀017562 in lung adenocarcinoma and normal lung tissue,and explore the possibility of hsa_circ₀017562 as a potential diagnostic marker of lung adenocarcinoma.2 To detect the influence of micro RNA miR-423-5p on proliferation,migration and invasion of lung adenocarcinoma cells and explore the possibility of miR-423-5p as a potential treatment of lung adenocarcinoma.Methods 1 Lung adenocarcinoma tissue and matched paracancerous normal lung tissue were collected during 2013/1²015/12 from 36 patients.The expression level of hsa_circ₀017562 in lung adenocarcinoma and normal lung tissue was detected by realtime PCR.Clinical significance of hsa_circ₀017562 expression was analyzed by the feature of clinicopathological in 36 patients.2 Culture A549,NCI-H1299,NCI-H157,A973 and GLC-82 lung adenocarcinoma cell lines,and the expression of miR-423-5p in the above cell lines was detected by real-time PCR.The miR-423-5p low expression cells were transiently transfected with miR-423-5p mimics to achieve over-expression.And miR-423-5p high expression cells were transiently transfected with miR-423-5p inhibitor to achieve knockdown.MTS and plate clone formation assay were used to detect the changes of cell proliferation ability.The changes of cell migration and invasion were observed by using transwell chamber.Results 1 The expression level of hsa_circ₀017562 in lung adenocarcinoma was significantly lower than that in normal lung tissue?4.82±0.55 VS 8.81±1.61,P<0.05?.There was no obvious difference in hsa_circ₀017562 expression level among the clinical stage,the smoking history and the pleural invasion.But significant difference was found in the gender group?P<0.05?.2 The expression level of miR-423-5p in A549,NCI-H1299,NCI-H157,A973 and GLC-82 cell lines was 0.92,0.88,0.22,0.19,0.47.The expression level of miR-423-5p in A549 cells transfected with miR-423-5p inhibitor was significantly lower than that in the control group?0.17±0.05 VS 0.91±0.06?,achieving knockdown effect.The expression level of miR-423-5p in A973 cells transfected with miR-423-5p mimics was significantly higher than that in the control group?1.22±0.13 VS 0.19 ± 0.07?,achieving over-expression effect.MTS:The proliferative activity of A549 cells with miR-423-5p inhibitor was significantly lower than that of the control group at 24 h,48h,72 h and 96 h.The proliferative activity of A973 cells with miR-423-5p mimics was significantly higher than that of control group at 24 h,48h,72 h and 96 h.The plate clone experiment showed the clone numbers of A549 cells with miR-423-5p inhibitor was lower than that of the control group?126 ± 28 VS 407 ± 23?;The clone numbers of A973 cells with miR-423-5p mimics was higher than that of the control group?535 ± 23 VS 385 ± 38?.Transwell migration experiments showed that the transmembrane cell numbers of A549 cell with miR-423-5p inhibitor was lower than that of the control group?89 ± 16 VS 143 ± 25?;however,the numbers of A973 cell with miR-423-5p mimics was higher than control group?223±30 VS 154 ± 35?.Transwell invation experiments showed that the transmembrane cell numbers of A549 cell with miR-423-5p inhibitor was lower than that of the control group?83±14 VS 117±20?,however,the numbers of A973 cell with miR-423-5p mimics was higher than control group?192±27 VS 133±28?.All the results were statistically significant?P<0.01?,suggesting that miR-423-5p plays a positive role in proliferation,migration and invasion of lung adenocarcinoma cells.Conclusions 1 The expression level of hsa_circ₀017562 in lung adenocarcinoma was significantly lower than that in normal lung tissue,which could be a potential molecular marker for the diagnosis of lung adenocarcinoma.2 miR-423-5p could positively regulate the proliferation,migration and invasion of lung adenocarcinoma cells,which could be a potential molecular target for the treatment of lung adenocarcinoma.