Targeting Cancer Progression Genes Upregulated in CREB3L1‐Deficient Breast Cancer Cells
Notice bibliographique
Résumé
Background Our laboratory discovered that CREB3L1 (cAMP responsive element binding protein 3‐like protein), a stress‐activated transcription factor, acts as a metastasis suppressor in breast cancer. CREB3L1 is expressed ubiquitously in non‐cancerous human breast cells and restricts expression of genes that promote cell growth, angiogenesis, and migration. Loss of CREB3L1 expression, which is a frequent event in high grade metastatic human breast tumors, can result in enhanced metastatic properties. To establish the mechanisms by which CREB3L1 exerts its tumor suppressive effects, a gene expression microarray analysis was performed to determine which genes are upregulated in CREB3L1‐deficient breast cancer cells. Six hundred and eighty genes were statistically significantly upregulated at least 2‐fold in all 4 of the CREB3L1‐deficient cell lines assessed. To focus our analysis, 4 genes were identified as potential targets for this study, given that they have previously been shown to have a role in cancer progression. Objective Gene expression of the 4 test genes (Genes 1, 2, 3, 4) are upregulated in CREB3L1‐deficient human breast cancer cells. Cells that had knockdown of each one of these 4 targets were extensively characterized to determine the role of each target in breast cancer tumorigenesis and metastasis. Hypothesis Inhibiting one or more of the cancer progression genes upregulated in CREB3L1‐deficient breast cancer cell lines will repress their tumorigenic and/or migratory properties. Methods Knockdowns were generated for each of the 4 genes in a CREB3L1‐deficient breast cancer cell line (HCC1806) and matched CREB3L1‐expressing cells (HCC1806 + HA‐CREB3L1), as well as a non‐tumorigenic control breast cell line (MCF10A). To knockdown expression of each of the 4 targets, sh RNA directed at the gene of interest was transduced via a lentiviral system into the cells. Cell‐based assays were performed to characterize the properties of these cell lines, which included the Boyden chamber assay to measure migration, soft agar assay to assess anchorage‐independent growth, and the MTT assay to evaluate cell proliferation. Results Successful knockdown at both the mRNA and protein level for each target gene was validated using quantitative real time PCR and immunoblotting. Results from the MTT assay verified that knockdown of each of the 4 gene targets did not affect cell survival in MCF10A cells, confirming that these test genes are not essential for survival in non‐cancerous breast cells (p > 0.05). Similarly, knockdown of each of the test genes did not affect cell proliferation in HCC1806 (± CREB3L1) cells, except for a significant decrease in proliferation in HCC1806 CREB3L1‐deficient cells with Gene 1 knockdown (p =0.02). Preliminary results indicate that knockdown of Genes 1, 2, and 4 in HCC1806 CREB3L1‐deficient cells reduces cancer cell properties, including cell migration and anchorage‐independent growth (p < 0.05). Conclusion Promising candidates will be tested in follow‐up mouse xenograft studies. Positive results in these pre‐clinical models would provide a strong rationale for the development and testing of inhibitors to one or more of these potential targets as a new therapeutic for the 30% of breast cancers that are CREB3L1‐deficient. Support or Funding Information This research received funding from Saskatchewan Health Research Foundation and the Canadian Institutes of Health Research.
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,001 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,002 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».