Interaction between p53 and Ras signaling controls cisplatin resistance via HDAC4- and HIF-1α-mediated regulation of apoptosis and autophagy

Abstract

The interplay between p53 and RAS signaling regulates cancer chemoresistance, but the detailed mechanism is unclear. In this study, we investigated the interactive effects of p53 and RAS on ovarian cancer cisplatin resistance to explore the potential therapeutic targets. Methods : An inducible p53 and RAS mutants active in either MAPK/ERK (S35 and E38) or PI3K/AKT (C40) or both (V12) were sequentially introduced into a p53-null ovarian cancer cell line-SKOV3. Comparative microarray analysis was performed using Gene Chip Prime View Human Gene Expression arrays (Affymetrix). In vitro assays of autophagy and apoptosis and in vivo animal experiments were performed by p53 induction and/or cisplatin treatment using the established cell lines. The correlation between HDAC4 and HIF-1α or CREBZF and the association of HDAC4, HIF-1α, CREBZF, ERK, AKT, and p53 mRNA levels with patient survival in 523 serous ovarian cancer cases from TCGA was assessed. Results : We show that p53 and RAS mutants differentially control cellular apoptosis and autophagy to inhibit or to promote chemoresistance through dysregulation of Bax, Bcl2, ATG3, and ATG12. ERK and AKT active RAS mutants are mutually suppressive to confer or to deprive cisplatin resistance. Further studies demonstrate that p53 induces HIF-1α degradation and HDAC4 cytoplasmic translocation and phosphorylation. S35, E38, and V12 but not C40 promote HDAC4 phosphorylation and its cytoplasmic translocation along with HIF-1α. Wild-type p53 expression in RAS mutant cells enhances HIF-1α turnover in ovarian and lung cancer cells. Autophagy and anti-apoptotic processes can be promoted by the overexpression and cytoplasmic translocation of HDAC4 and HIF1-α. Moreover, the phosphorylation and cytoplasmic translocation of HDAC4 activate the transcription factor CREBZF to promote ATG3 transcription. High HDAC4 or CREBZF expression predicted poor overall survival (OS) and/or progression-free survival (PFS) in ovarian cancer patients, whereas high HIF-1α expression was statistically correlated with poor or good OS depending on p53 status. Conclusion : HIF-1α and HDAC4 may mediate the interaction between p53 and RAS signaling to actively control ovarian cancer cisplatin resistance through dysregulation of apoptosis and autophagy. Targeting HDAC4, HIF-1α and CREBZF may be considered in treatment of ovarian cancer with p53 and RAS mutations.

The crosstalk between STAT3 and p53/RAS signaling controls cancer cell metastasis and cisplatin resistance via the Slug/MAPK/PI3K/AKT-mediated regulation of EMT and autophagy

“…Because p53 is commonly mutated and PI3K/AKT and MAPK-associated molecuels belong to RAS signaling 20,21 , we wondered if p53 and RAS are invovled in the STAT3-mediated autophagy, tumor growth, and cisplatin resistance. To investigate the effects of RAS and p53 on STAT3 function, we first introduced a dox-induction system to induce p53 expression in SKOV3 cells deficient with wild type p53, and then added a RAS mutant V12 active with both MAPK and PI3K/AKT into these cells afterwards 15 . The cDNA of STAT3-DN was also introduced into above cell lines (Fig.…”

“…To establish SKOV3 T cell lines expressing HRAS-V12, the full length RAS V12 cDNA was amplified from previously established plasmids in our lab using the primers 5′-CGCggatccATGACCGAATACAAGCTTGTTG -3′ (forward; lower case letters represent the Bam HI site) and 5′-TGAT ctcgag TCAatggtgatggtgatgatgGGAGAGCACACACTTGCAGCTCA-3′ (reverse; italic lower case letters represent the Xho I site; bold lower case letters indicate the His tag), digested with Bam HI and Xho I, and inserted into the retrovirus vector pBabe-zeo. The correct plasmids were confirmed by sequencing 15 . Constitutive activated STAT3 (STAT3-C), wild-type STAT3 (STAT3-WT), and dominant negative STAT3 (STAT3-DN) containing HA-tag plasmids were constructed into lentiviral vector pCDH-CMV-MCS-EF1- puro with Eco RI and Bam HI from the original plasmids purchased from Addgene.…”

Characterization of an Amphiphilic Phosphonated Calixarene Carrier Loaded With Carboplatin and Paclitaxel: A Preliminary Study to Treat Colon Cancer in vitro and in vivo

“…Its naturally aggressive progression, in combination with a lack of accurate screening and detection methods, means that patients are typically diagnosed at advanced stages, and they therefore respond inadequately to available treatments (Blind et al, 2018). Chemotherapies are essential due to the high risk of relapse after surgery, yet most drugs for treating advanced-stage colorectal cancer, such as cisplatin, are relatively ineffective and frequently induce adverse side effects (Rabik and Dolan, 2007; Robella et al, 2019), such as nephro-(Zhu et al, 2019), hepato-(Zhang X. et al, 2019), and cardiotoxicity (Dasari and Tchounwou, 2014). These considerations highlight the need for new therapeutic approaches for this disease.…”

MUC3A Promotes Non-Small Cell Lung Cancer Progress via Activating NFκB Pathway and Attenuates Radiosensitivity

“…It has been reported that NFκB signaling induces angiogenesis by increasing the production of VEGF [44][45][46], angiogenesis induced by MUC3A is also mediated by NFκB activation in NSCLC. Radiation-mediate hypoxia triggers HIF-1α transcription and up-regulates VEGF expression [46,47], MUC3A knockdown may enhance radiosensitivity by promoting oxygen stress and impair HIF-1α and VEGF expression level. Furthermore, MMP-2 and MMP-9 are also known to stimulate tumor angiogenesis and the epithelial-to-mesenchymal transition (EMT) through partial proteolysis of the ECM [48][49][50][51][52].…”