Abstract
Lung cancer is a leading cause of cancer death all around the world. Long non-coding RNAs (lncRNAs) have been confirmed to be involved in carcinogenesis of malignancies. However, the molecular mechanism of most lncRNAs in various kinds of cancers remains unclear. LncRNA HOTAIR and HNRNPA1 are reported to play an oncogenic role in non-small cell lung cancer, and the overexpression of HNRNPA1 is shown to promote the proliferation of lung adenocarcinoma cells. In our study, we find that the overexpression of HOTAIR could promote the proliferation and overexpression of miR-149-5p could inhibit the proliferation of lung cancer cells. Flow cytometric analysis determines that overexpression of miR-149-5p induces cell cycle arrest in the G0/G1 phases, whereas overexpression of HOTAIR decreases the proportion of G0/G1phase cells. Also, overexpression of HOTAIR promotes the migration and invasion ability of lung cancer cells, confirmed by the wound-healing and transwell assays, which are suppressed by overexpression of miR-149-5p. Furthermore, the dual-luciferase reporter assay indicates that miR-149-5p could bind both HOTAIR and the 3′UTR of HNRNPA1. In summary, we find that HOTAIR can regulate HNRNPA1 expression through a ceRNA mechanism by sequester miR-149-5p, which post-transcriptionally targets HNRNPA1, thus promoting lung cancer progression.
Highlights
Lung cancer is the most lethal and frequently diagnosed malignant tumor
We find that HOTAIR may act as a competing endogenous RNAs for miR-149-5p to upregulate the expression of HNRNPA1
The results show that a significant reduction in luciferase reporter activity was observed in group of cells co-transfected with pmir-h-HOTAIR-WT and miR-149-5p mimics and group of cells co-transfected with pmir-h-HNRNPA1-WT and miR-1495p mimics (Figure 4C)
Summary
Lung cancer is the most lethal and frequently diagnosed malignant tumor. Approximately 2.09 million new cases were diagnosed, and 1.76 million died of lung cancer around the world in 2018 [1]. Expression of miR-149-5p and HOTAIR in Lung Cancer Cell Lines. The background expression level of miR-149-5p and HOTAIR in A549, SPC-A-1, and HBE cell lines were quantified by qRTPCR (Figure 1B). The expression of miR-149-5p was lower in the SPC-A-1 cell line compared to that in A549 and HBE, and SPC-A-1 was chosen to perform miR-149-5p overexpressed lentivirus transfection, and the overexpression level of miR-149-5p was detected by qRTPCR (Figure 1D). Background expression of HOTAIR in the SPC-A-1 cell line was higher than that in HBE cell line; siHOTAIR and siNC were transfected to SPCA-1 cell line to knock down HOTAIR expression to perform the gain- and loss-of-function experiments for investigating the effect of HOTAIR on lung cancer cells. The knockdown efficiency of HOTAIR was detected and quantified by qRT-PCR, and the result showed that siHOT-1597 had the highest knockdown efficiency (Figure 1F)
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