Screening for alternative splicing of lncRNA Dleu2 in the mouse liver cell line AML-12

Abstract

The long non coding RNA deleted in leukemia 2 gene (Dleu2) has recently been demonstrated to be an active player in the progression of several types of cancer, including hepatocellular carcinoma. Dleu2 may serve a role in modulating the downstream effects-mediated by alternative splicing of its multiple exons. However, the proportional expression of these alternative splicing populations of the Dleu2 exons is currently unknown. To determine how Dleu2 could be affected by alternative splicing, a series of alternative splicing primer sets were designed to investigate which transcripts were preferentially activated when Dleu2 was targeted for downregulation or upregulation. A specific Dleu2 small interfering RNA that targeted an exon upstream of the tumor suppressor microRNA site significantly knocked down Dleu2 expression across all the primer sets used, which targeted 13 different alternative splicing transcripts over 5 different promoter sites in the mouse liver cell line, AML-12. Similarly, 50 µM Resveratrol led to significant upregulation of Dleu2 in 11 alternative splicing transcripts. These results show that Dleu2 is capable of successful modulation across alternative splicing transcripts that can be screened, and also that Resveratrol can be a potential nutraceutical, which may potentially lead to novel approaches in the use of lncRNA Dleu2 for diagnostics and regulation.

Contribution of Non-Coding RNAs to Anticancer Effects of Dietary Polyphenols: Chlorogenic Acid, Curcumin, Epigallocatechin-3-Gallate, Genistein, Quercetin and Resveratrol

“…Kay et al [ 194 ] demonstrated that RES upregulated the tumor suppressor gene DLEU2 in 11 alternative splicing transcripts. Since DLEU2 was shown to negatively regulate cyclins E1 and D1 through upregulation of miR-15a/miR-16-1 and since overexpression of DLEU2 recovered cellular proliferation and inhibition of the colony-forming ability of tumor cells in a miR-15a/miR-16-1-dependent manner [ 195 ], RES’s upregulation of DLEU2 may contribute to its anticancer effects.…”