LINC00342调控miR-384对肺鳞癌细胞增殖侵袭迁移的影响研究

LINC00342 Regulates the Effect of miR-384 on the Proliferation Invasionand Migration of Lung Squamous Carcinoma Cells

目的:探讨LINC00342通过miR-384对肺鳞癌细胞增殖、侵袭、迁移的影响。方法:将肺鳞癌SK-MES-1细胞分为:ctrl组(正常培养的SK-MES-1细胞)、si-NC组(转染si-NC)、si-LINC00342组(转染si-LINC00342)、si-LINC00342+inhibitor-NC组(si-LINC00342和inhibitor-NC共转染)、si-LINC00342+miR-384 inhibitor组(si-LINC00342和miR-384 inhibitor共转染);RT-qPCR检测细胞中LINC00342、miR-384表达;CCK-8法及Transwell小室法分别检测细胞增殖和侵袭;划痕实验及流式细胞仪分别检测细胞迁移和凋亡;Western blot检测细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)、MMP-9、半胱氨酸蛋白酶3(caspase-3)的表达;双荧光素酶报告基因实验验证LINC00342和miR-384的关系。结果:与ctrl组、si-NC组比较,si-LINC00342组SK-MES-1细胞的OD450值、细胞侵袭数目、划痕愈合率、PCNA、MMP-2和MMP-9表达明显降低,细胞凋亡率、caspase-3表达明显上升(P<0.05);抑制miR-384表达减弱了敲低LINC00342对SK-MES-1细胞增殖、侵袭、迁移的抑制作用,降低了细胞凋亡能力;LINC00342靶向调控miR-384表达。结论:敲低LINC00342可能通过靶向miR-384,进而抑制SK-MES-1细胞增殖、侵袭和迁移。

Objective:To investigate the influences of LINC00342 on the proliferation,invasion,and migration of lung squamous cell carcinoma through miR-384.Methods:Lung squamous cell carcinoma SK-MES-1 cells were grouped into the ctrl group(normally cultured SK-MES-1 cells),si-NC group(transfected with si-NC),and si-LINC00342 group(transfected with si-LINC00342),si-LINC00342+inhibitor-NC group(co-transfected with si-LINC00342 and inhibitor-NC),and si-LINC00342+miR-384 inhibitor group(co-transfected with si-LINC00342 and miR-384 inhibitor);RT-qPCR was applied to detect the expression of LINC00342 and miR-384 in cells;CCK-8 assay and Transwell chamber were applied to detect cell proliferation and invasion;scratch assay and flow cytometry were used to cell migration and apoptosis;Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),MMP-9,and cysteine protease-3(caspase-3)in cells;and dual-luciferase reporter assay was used to verify the relationship between LINC00342 and miR-384.Results:Compared with the ctrl group and si-NC group,the OD450 value,the number of cell invasions,the wound healing rate,the expressions of PCNA,MMP-2,and MMP-9 in SK-MES-1 cells in the si-LINC00342 group was obviously decreased,and the cell apoptosis was obviously decreased,the mortality rate and the expression of caspase-3 were obviously increased(P<0.05);inhibiting the expression of miR-384 attenuated the inhibitory effect of knockdown of LINC00342 on the proliferation,invasion,and migration of SK-MES-1 cells,and reduced the apoptosis ability;LINC00342 targeted miR-384 expression.Conclusion:Knockdown of LINC00342 may inhibit the proliferation,invasion,and migration of SK-MES-1 cells by targeting miR-384.