RÉSUMÉ
Although ALK tyrosine kinase inhibitors (ALK-TKIs) have shown remarkable benefits in EML4-ALK positive NSCLC patients compared to conventional chemotherapy, the optimal sequence of ALK-TKIs treatment remains unclear due to the emergence of primary and acquired resistance and the lack of potential prognostic biomarkers. In this study, we systematically explored the validity of sequential ALK inhibitors (alectinib, lorlatinib, crizotinib, ceritinib and brigatinib) for a heavy-treated patient with EML4-ALK fusion via developing an in vitro and in vivo drug testing system based on patient-derived models. Based on the patient-derived models and clinical responses of the patient, we found that crizotinib might inhibit proliferation of EML4-ALK positive tumors resistant to alectinib and lorlatinib. In addition, NSCLC patients harboring the G1269A mutation, which was identified in alectinib, lorlatinib and crizotinib-resistant NSCLC, showed responsiveness to brigatinib and ceritinib. Transcriptomic analysis revealed that brigatinib suppressed the activation of multiple inflammatory signaling pathways, potentially contributing to its anti-tumor activity. Moreover, we constructed a prognostic model based on the expression of IL6, CXCL1, and CXCL5, providing novel perspectives for predicting prognosis in EML4-ALK positive NSCLC patients. In summary, our results delineate clinical responses of sequential ALK-TKIs treatments and provide insights into the mechanisms underlying the superior effects of brigatinib in patients harboring ALKG1269A mutation and resistant towards alectinib, lorlatinib and crizotinib. The molecular signatures model based on the combination of IL6, CXCL1 and CXCL5 has the potential to predict prognosis of EML4-ALK positive NSCLC patients.
Sujet(s)
Adénocarcinome pulmonaire , Antinéoplasiques , Tumeurs du poumon , Protéines de fusion oncogènes , Composés organiques du phosphore , Inhibiteurs de protéines kinases , Pyrimidines , Humains , Composés organiques du phosphore/usage thérapeutique , Composés organiques du phosphore/pharmacologie , Pyrimidines/usage thérapeutique , Pyrimidines/pharmacologie , Tumeurs du poumon/traitement médicamenteux , Tumeurs du poumon/génétique , Tumeurs du poumon/anatomopathologie , Protéines de fusion oncogènes/génétique , Protéines de fusion oncogènes/métabolisme , Animaux , Adénocarcinome pulmonaire/traitement médicamenteux , Adénocarcinome pulmonaire/génétique , Adénocarcinome pulmonaire/anatomopathologie , Inhibiteurs de protéines kinases/usage thérapeutique , Inhibiteurs de protéines kinases/pharmacologie , Pronostic , Antinéoplasiques/usage thérapeutique , Antinéoplasiques/pharmacologie , Résistance aux médicaments antinéoplasiques , Lactames/usage thérapeutique , Carbazoles/usage thérapeutique , Carbazoles/pharmacologie , Sulfones/usage thérapeutique , Sulfones/pharmacologie , Crizotinib/usage thérapeutique , Crizotinib/pharmacologie , Lignée cellulaire tumorale , Pipéridines/usage thérapeutique , Pipéridines/pharmacologie , Femelle , Souris , Inflammation/traitement médicamenteux , Carcinome pulmonaire non à petites cellules/traitement médicamenteux , Carcinome pulmonaire non à petites cellules/génétique , Pyrazoles/usage thérapeutique , Pyrazoles/pharmacologie , Mâle , Kinase du lymphome anaplasique/génétique , Kinase du lymphome anaplasique/antagonistes et inhibiteurs , Kinase du lymphome anaplasique/métabolisme , Prolifération cellulaire/effets des médicaments et des substances chimiques , Mutation , Aminopyridines/usage thérapeutique , Aminopyridines/pharmacologieRÉSUMÉ
BACKGROUND: Metastasis accounts for the most lethal reason for the death of ovarian cancer patients, but remains largely untreated. Epithelial-mesenchymal transition (EMT) is critical for the conversion of early-stage ovarian tumours into metastatic malignancies. Thus the exploration of the signalling pathways promoting EMT would open potential opportunities for the treatment of metastatic ovarian cancer. Herein, the putative role of MDM2 in regulating EMT and metastasis of ovarian cancer SKOV3 cells was investigated. METHODS: The regulatory effects by MDM2 on cell motility was emulated by wound-healing and transwell assays. The effects on EMT transition and Smad pathway were studied by depicting the expression levels of epithelial marker E-cadherin as well as key components of Smad pathway. To evaluate the clinical relevance of our findings, the correlation of MDM2 expression levels with the stages of 104 ovarian cancer patients was investigated by immunohistochemistry assay. RESULTS: We demonstrate that MDM2 functions as a key factor to drive EMT and motility of ovarian SKOV3 cells, by facilitating the activation of TGF-ß-Smad pathway, which results in the increased transcription of snail/slug and the subsequent loss of E-cadherin levels. Such induction of EMT is sustained in either E3 ligase-depleted MDM2 or E3 ligase inhibitor HLI-373-treated cells, while being impaired by the N-terminal deletion of MDM2, which is also reflected by the inhibitory effects against EMT by Nutlin-3a, the N-terminal targeting agent. The expression levels of MDM2 is highly correlated with the stages of the ovarian cancer patients, and the higher expression of MDM2 together with TGFB are closely correlated with poor prognosis and predict a high risk of ovarian cancer patients. CONCLUSIONS: This study suggests that MDM2 activates Smad pathway to promote EMT in ovarian cancer metastasis, and targeting the N-terminal of MDM2 can reprogram EMT and impede the mobility of cancer cells.
Sujet(s)
Carcinomes/génétique , Transition épithélio-mésenchymateuse/génétique , Tumeurs de l'ovaire/génétique , Protéines proto-oncogènes c-mdm2/génétique , Aminoquinoléines/pharmacologie , Antigènes CD , Technique de Western , Cadhérines/métabolisme , Carcinomes/métabolisme , Carcinomes/anatomopathologie , Lignée cellulaire tumorale , Mouvement cellulaire/génétique , Transition épithélio-mésenchymateuse/effets des médicaments et des substances chimiques , Femelle , Technique d'immunofluorescence , Techniques de knock-down de gènes , Humains , Immunohistochimie , Métastase tumorale , Stadification tumorale , Tumeurs de l'ovaire/métabolisme , Tumeurs de l'ovaire/anatomopathologie , Protéines proto-oncogènes c-mdm2/effets des médicaments et des substances chimiques , Protéines proto-oncogènes c-mdm2/métabolisme , Réaction de polymérisation en chaine en temps réel , Transduction du signal , Protéines Smad/métabolisme , Facteurs de transcription de la famille Snail/génétique , Thymine/analogues et dérivés , Thymine/pharmacologie , Facteur de croissance transformant bêta/métabolismeRÉSUMÉ
Although hypoxia is a prominent feature contributing to the therapeutic resistance of hepatocellular carcinoma cells (HCC) against chemotherapeutic agents, including the Topoisomerase I inhibitor SN38, the underlying mechanism is not fully understood and its understanding remains a major clinical challenge. In the present study, we found that hypoxia-induced nuclear translocation and accumulation of YAP acted as a survival input to promote resistance to SN38 in HCC. The induction of YAP by hypoxia was not mediated by HIF-1α because manipulating the abundance of HIF-1α with CoCl2, exogenous expression, and RNA interference had no effect on the phosphorylation or total levels of YAP. The mevalonate-HMG-CoA reductase (HMGCR) pathway may modulate the YAP activation under hypoxia. Combined YAP inhibition using either siRNA or the HMGCR inhibitor statins together with SN38 treatment produced improved anti-cancer effects in HCC cells. The increased anti-cancer effect of the combined treatment with statins and irinotecan (the prodrug of SN-38) was further validated in a human HepG2 xenograft model of HCC in nude mice. Taken together, our findings identify YAP as a novel mediator of hypoxic-resistance to SN38. These results suggest that the administration of SN28 together with the suppression of YAP using statins is a promising strategy for enhancing the treatment response in HCC patients, particularly in advanced stage HCC cases presenting hypoxic resistance.