MiR-489-3p Affect The Proliferation Of Lung Cancer Cells By Regulating MCM2

ObjectivemiR-489-3p can be used as a targeted therapeutic target,down-regulated in a variety of tumor cells,over expression can inhibit tumor growth to a certain extent,it is a kind of tumor suppressor.In this paper,we will confirm that mir-489-3p may affect the proliferation of lung cancer cells by regulating MCM2.MethodsIn this study,A549 lung cancer cells and BEAS-2B normal lung epithelial cells were purchased from Shanghai cell bank of Chinese Academy of Sciences.After inoculation and culture,the role and regulatory mechanism of mir-489-3p in the development of lung cancer were explored.Mir-489-3p sequence was obtained by miRBase database,mir-489-3pmimics was designed and transfected into A549 cells.The effect of over expression of mir-489-3p on the proliferation of A549 cells was detected by CCK8 experiment,and the regulatory protein MCM2 was detected by real-time fluorescent quantitative PCR and Western blotting.Finally,the target relationship between mir-489-3p and MCM2 was confirmed by double luciferase reporter gene experiment.ResultsThe results showed that the expression level of mir-489-3p and MCM2 mRNA in A549,H1299 and SPC-A1 cells was significantly lower than that in BEAS-2B cells;the expression level of MCM2 protein in A549,H1299 and SPC-A1 cells was significantly higher;the expression level of Bcl-2 protein in A549 cells in mir-489-3p group was significantly lower than that in normal lung epithelial cells The expression level of Caspase-3 and Bax protein was significantly increased;the apoptosis rate of A549 cells in mir-489-3p group was significantly increased;the expression level of MCM2,CyclinD1 and bcl-2 protein in A549 cells in Si MCM2 group was significantly decreased,and the expression level of p21 and Bax protein was significantly increased;the activity of A549 cells in Si MCM2 group was 48 h and 72 h respectively The apoptosis rate of A549 cells in the si-mcm2 group was significantly higher than that in the control group.The luciferase activity of wt-mcm2 cells in the mir-489-3p group was significantly lower than that in the mir-489-3p group.The luciferase activity of the mut-mcm2 cells in the mir-489-3p group was not significantly different from that in the mir-489-3p group The expression level of MCM2,CyclinD1,Bcl-2 and p21,Bax in A549 cells of mir-489-3p group were significantly lower than that of anti-mir-489-3p group The activity of A549 cells in mir-489-3p group decreased significantly at 48 h and 72 h;the activity of A549 cells in mir-489-3p+pcdna-mcm2 group increased significantly at 48 h and 72 h;the apoptosis rate of A549 cells in mir-489-3p group increased significantly;the apoptosis rate of A549 cells in mir-489-3p+pcdna-mcm2 group decreased significantly.ConclusionsThe data showed that over expression of mir-489-3p could significantly inhibit the proliferation of A549 cells.Mir-489-3p can inhibit the proliferation and promote the apoptosis of lung cancer A549 cells,which is closely related to the targeted regulation of MCM2,which will provide a new direction and a new way for cancer prevention and treatment.