linc00619通过AKT/ERK信号抑制BEAS-2B细胞的增殖和迁移

Linc00619 gene inhibits the proliferation and migration of BEAS-2B cells through AKT/ERK signaling

目的:探讨长链非编码RNA 00619(linc00619)在人肺上皮细胞BEAS-2B中的生物学作用。方法:用RNA原位杂交对linc00619进行BEAS-2B亚细胞定位;采用CRISPR/Cas9方法构建linc00619慢病毒敲除载体,CCK8和Transwell试验检测linc00619敲除对BEAS-2B增殖和迁移的影响;lncRNA芯片检测linc00619敲除后差异表达的mRNAs并进行KEGG富集分析;Western blot检测AKT、MEK和ERK等增殖相关分子。结果:linc00619定位于细胞质。linc00619敲除促进了BEAS-2B的增殖和迁移。linc00619敲除后得到了3461个差异表达mRNAs。共有922个mRNAs被富集进KEGG信号通路,其中MAPK信号是显著富集的信号之一。Western blot证实,linc00619敲除活化了AKT和ERK。结论:细胞质表达的linc00619通过抑制AKT/ERK信号抑制了BEAS-2B的增殖和迁移。

Objective:To investigate the biological role of linc00619 gene in human lung epithelial cells.Methods:Linc00619 gene in BEAS-2B cells was subjected to subcellular localization using RNA in situ hybridization,and linc00619-knockout lentivirus was constructed using CRISPR/Cas9 technique.Then,the effect of linc00619 knockout on the BEAS-2B cell was observed through cell proliferation and invasion assays.LncRNA microarray was used to detect the differentially expressed mRNAs following linc00619 knockout,and KEGG signaling pathway was also enriched.The proliferation-related molecules,AKT,MEK and ERK were determined by Western blot.Results:Linc00619 was localized in the cytoplasm,and LINC00619 knockout promoted the proliferation and invasion of BEAS-2B cells.Linc00619 knockout resulted in 3461 differentially expressed mRNAs after microarray screening.A total of 922 genes were enriched into the KEGG signaling pathway,in which MAPK signaling pathway was one of the significant enrichments,and AKT and ERK were activated by linc00619 knockout confirmed with Western blot.Conclusion:Highly expressed linc00619 in the cytoplasm may inhibit the proliferation and invasion of BEAS-2B cells via checking AKT/ERK signaling.