21045158

pubmed:Citation

22

Increasing evidence points to the functional importance of alternative splice variations in cancer pathophysiology. Two splice variants are derived from the CASP9 gene via the inclusion (Casp9a) or exclusion (Casp9b) of a four-exon cassette. Here we show that alternative splicing of Casp9 is dysregulated in non-small cell lung cancers (NSCLC) regardless of their pathologic classification. Based on these findings we hypothesized that survival pathways activated by oncogenic mutation regulated this mechanism. In contrast to K-RasV12 expression, epidermal growth factor receptor (EGFR) overexpression or mutation dramatically lowered the Casp9a/9b splice isoform ratio. Moreover, Casp9b downregulation blocked the ability of EGFR mutations to induce anchorage-independent growth. Furthermore, Casp9b expression blocked inhibition of clonogenic colony formation by erlotinib. Interrogation of oncogenic signaling pathways showed that inhibition of phosphoinositide 3-kinase or Akt dramatically increased the Casp9a/9b ratio in NSCLC cells. Finally, Akt was found to mediate exclusion of the exon 3,4,5,6 cassette of Casp9 via the phosphorylation state of the RNA splicing factor SRp30a via serines 199, 201, 227, and 234. Taken together, our findings show that oncogenic factors activating the phosphoinositide 3-kinase/Akt pathway can regulate alternative splicing of Casp9 via a coordinated mechanism involving the phosphorylation of SRp30a.

eng

IM

MEDLINE

1538-7445

Copyright © 2010 AACR.

15

NLM

9185-96

pubmed-meshheading:21045158-Alternative Splicing, pubmed-meshheading:21045158-Blotting, Western, pubmed-meshheading:21045158-Carcinoma, Non-Small-Cell Lung, pubmed-meshheading:21045158-Caspase 9, pubmed-meshheading:21045158-Cell Line, Tumor, pubmed-meshheading:21045158-Cell Proliferation, pubmed-meshheading:21045158-Gene Expression Regulation, Enzymologic, pubmed-meshheading:21045158-Gene Expression Regulation, Neoplastic, pubmed-meshheading:21045158-Humans, pubmed-meshheading:21045158-Isoenzymes, pubmed-meshheading:21045158-Lung Neoplasms, pubmed-meshheading:21045158-Mutation, pubmed-meshheading:21045158-Nuclear Proteins, pubmed-meshheading:21045158-Phosphatidylinositol 3-Kinases, pubmed-meshheading:21045158-Phosphorylation, pubmed-meshheading:21045158-Protein Kinase Inhibitors, pubmed-meshheading:21045158-Proto-Oncogene Proteins c-akt, pubmed-meshheading:21045158-Quinazolines, pubmed-meshheading:21045158-RNA, Messenger, pubmed-meshheading:21045158-RNA Interference, pubmed-meshheading:21045158-RNA-Binding Proteins, pubmed-meshheading:21045158-Receptor, Epidermal Growth Factor, pubmed-meshheading:21045158-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:21045158-Signal Transduction

Alternative splicing of caspase 9 is modulated by the phosphoinositide 3-kinase/Akt pathway via phosphorylation of SRp30a.

Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, N.I.H., Extramural