Alternate Splicing Factor Fox2 Affects Growth and Survival of Multiple Myeloma Cells
Bibliographic record
Abstract
Abstract Abstract 1810 Alternative splicing of pre-mRNA plays a critical role in number of cellular processes including cell growth and survival. We have previously demonstrated significant frequency of alternate splicing in multiple myeloma (MM) that points to the need for evaluation of not only the expression level of genes but also post translational modifications. Our data utilizing exon array profile from 170 uniformly treated newly diagnosed patients with MM confirms clinical relevance of splicing as demonstrated by impact of level and extent of alternate splicing on both progression free and overall survival. We have now further studied the molecular mechanisms that may be involved in differential splicing in myeloma. We have evaluated the expression data from 170 uniformly treated newly diagnosed patients with MM and identified Fox-2, an RNA binding protein that is thought to be a key regulator of alternative exon splicing as one of the most important gene predicting clinical outcome in MM. We have now investigated the molecular effects of Fox2 and its role on myeloma cell growth and survival. Using Fox-2-directed shRNA lentivirus, we knocked-out (KO) the expression of Fox2 in MM1S and RPMI8226 cell lines. Real-time PCR and western blot analysis confirmed KO of over 90% of Fox2 expression in both cell lines. We observed significant inhibition of proliferation of both MM1S and RPMI8226 cell lines in Fox-2- KO compared to control cell lines. Using annexin V and PI staining and flow cytometric analysis we also observed induction of apoptosis in Fox-2-KO-MM1S and RPMI8226 cells. The apoptosis was Caspase 3 and caspase 8-mediated in the Fox-2-KO. A similar effect of Fox-2-KO was observed on cell survival and proliferation in Hela and 293 cells. Fox-2 also plays a crucial role in the maintenance of cytoskeleton of MM cells. We observed that actin was significantly downregulated in both MM1S and RPMI8226 knockdown cell lines compare to controls, whereas GAPDH expressions are equal in all conditions. It's been recognized that actin derangement is associated with cell apoptosis. The RNA binding specificity of Fox-2 in MM cells by CLIP-seq and changes in alternate splicing in Fox-2-KO by exon array is underway to understand the molecular events mediated by this transcriptome modifier affecting MM pathobiology. These results identify Fox-2 as a biologically important intermediate with essential function in myeloma with potential biological and clinical implications. Disclosures: No relevant conflicts of interest to declare.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".