Effect Of LncRNA GAS5 On Cisplatin Resistance In Lung Adenocarcinoma

Objective:To explore the influence and regulation mechanism of long non-coding RNA(lnc RNA)growth arrest-specific transcription factor 5(GAS 5)on cisplatin(DDP)resistance in lung cancer cells.Methods:(1)A549 and A549 / DDP cells were divided into three groups: Control,LV-sg-NC and LV-sg-Lnc RNA GAS5.48 h After plasmid transfection,another drug treatment(DDP 30μg / m L)was treated for 24 h.The lnc RNA GAS5 expression level of A549 and A549 / DDP cells after sg-Lnc RNA GAS5 virus was quantitatively determined by q RT-PCR experimental technology.(2)The effect of A549,A549 / DDP cells transfected and cisplatin was measured by CCK-8 method.The cells were divided into three groups,Control,sg-NC and sg-Lnc RNA GAS5.The cells were treated with cisplatin for 24 h and cisplatin drug concentration: 0,10,20,30,40,50 and 60 μ g / m L.Cell activity and IC50 were calculated based on OD to explore the change in sensitivity of A549,A549 / DDP cells after transfection with sh-lnc RNA GAS5.(3)A549 and A549 / DDP cells were transfected with liposome to construct lnc RNA GAS5 overexpression and low expression models,which were divided into Control,OE-NC,OE-Lnc RNA GAS5,sg-NC and sg-Lnc RNA GAS5.The cells to be transfected were stable and treated with cisplatin.The effect of lung cancer cell proliferation ability was observed by plate clone formation experiment.The number of clonal cells was counted,and the colonies of 50 cells were observed under the microscope.(4)detecting the targeting interaction between lnc RNA GAS5 and mi R-152-3p mimics by bioinformatics prediction and dual-luciferase assay,The dual-luciferase assay performed with 293 T cells,They were divided into groups Control,GAS 5-WT + mimics NC,GAS 5-WT + mi R-152-3p mimics,GAS 5-Mut + mimics NC,and GAS 5-Mut + mi R-152-3p mimics,After 48 h of transfection,Luciferase activity in each group was measured separately by dual luciferase assay,Based on the test data,the two luciferase ratios(i.e.reporter gene / reference gene),Comparing ratio differences between different groups.Results:(1)According to q RT-PCR results,sg-Lnc RNA GAS5 infection in A549 and A549 / DDP cells(P <0.001),and the transfection was successful.(2)After transfection of sg-Lnc RNA GAS5 in A549 and A549 / DDP cells,the cell survival rate of sg-Lnc RNA GAS5 group compared with sg-NC group(P <0.001,significant difference),which promoted cell proliferation and cisplatin resistance.(3)In the A549 cells,OE-Lnc RNA GAS5 compared with OE-NC group(P <0.001),Inhibit cell proliferation;Comparing the sg-Lnc RNA GAS5 with the sg-NC group,Significant increase in clone formation(P <0.001),Promote cell proliferation;In the A549 / DDP cells,The OE-Lnc RNA GAS5,when compared with the OE-NC group,Significant decrease in the number of clone formation(P <0.001),Inhibit cell proliferation;Comparing the sg-Lnc RNA GAS5 with the sg-NC group,Significant increase in clone formation(P <0.001),And promote cell proliferation.(4)According to the dual luciferase assay,the relative fluorescence intensity of GAS 5-WT + mi R-152-3p mimics group(15.80 ± 0.26)decreased significantly compared with the GAS 5-WT + mimics NC group(19.42 ± 0.38)(P <0.001),This result suggested that mi R-152-3p can combine with the gene sequence of GAS 5 to attenuate the expression level of GAS 5 gene.Conclusion:Low expression of lnc RNA GAS5 promoted proliferation and cisplatin resistance in lung cancer cells;high lnc RNA GAS5 expression inhibited lung cancer cell proliferation and reduced cisplatin resistance in lung cancer;there is a targeting relationship between lnc RNA GAS5 and mi R-152-3p,and lnc RNA GAS5 may be a target gene directly regulated by mi R-152-3p.The lnc RNA GAS5 gene may be a valid potential target for reversing cisplatin chemosistance in human non-small cell lung cancer.