A prototypical non-malignant epithelial model to study genome dynamics and concurrently monitor micro-RNAs and proteins in situ during oncogene-induced senescence

摘要: Senescence is a fundamental biological process implicated in various pathologies, including cancer. Regarding carcinogenesis, senescence signifies, at least its initial phases, an anti-tumor response that needs to be circumvented for cancer progress. Micro-RNAs, subclass of regulatory, non-coding RNAs, participate regulation. At the subcellular level micro-RNAs, similar proteins, have been shown traffic between organelles influencing cellular behavior. The differential function micro-RNAs relative their localization and role biology raises concurrent situ analysis coding gene products senescent cells as necessity. However, technical challenges rendered co-detection unfeasible until now. In present report we describe methodology bypasses these limitations achieving first time simultaneous detection both micro-RNA protein context senescence, utilizing new commercially available SenTraGorTM compound. method was applied prototypical human non-malignant epithelial model oncogene-induced generated purposes study. For characterization this novel system, wide range molecular techniques, well high-throughput transcriptome micro-RNAs. This experimental setting has three advantages are presented discussed: i) it covers “gap” carcinogenesis field, almost all corresponding vitro models fibroblast-based, even though majority neoplasms origin, ii) recapitulates precancerous cancerous phases tumorigenesis within short frame under light natural selection iii) uses oncogenic signal, replication licensing factor CDC6, DNA transcription when over-expressed, characteristic can exploited monitor RNA dynamics. Consequently, demonstrate our optimal studying basis shedding on tumor-initiating events. latter may reveal targets with clinical benefit. Besides, since incorporated low, medium or image-based approaches, expect broadly applicable.