Skp2-Mediated Stabilization of MTH1 Promotes Survival of Melanoma Cells upon Oxidative Stress

Wang, Jia Yu; Liu, Guang Zhi; Wilmott, James S.; La, Ting; Feng, Yu Chen; Yari, Hamed; Yan, Xu Guang; Thorne, Rick F.; Scolyer, Richard A.; Zhang, Xu Dong; Jin, Lei

doi:10.1158/0008-5472.can-17-1965

Cited by 44 publications

(28 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The human melanoma cell lines Mel-RM, Sk-Mel-28, A375, and mouse melanoma cell line B16 were cultured in DMEM containing 5% FCS (14). HEK293T, A549, MCF7, HeLa, WiDr, HT29, HCT116, U2OS cells were obtained from ATCC and cultured according to ATCC instructions.…”

Section: Cell Culturementioning

confidence: 99%

See 1 more Smart Citation

A p53-Responsive miRNA Network Promotes Cancer Cell Quiescence

Liu

Farrelly

et al. 2018

Cancer Research

Self Cite

View full text Add to dashboard Cite

Cancer cells in quiescence (G 0 phase) are resistant to death, and re-entry of quiescent cancer cells into the cellcycle plays an important role in cancer recurrence. Here we show that two p53-responsive miRNAs utilize distinct but complementary mechanisms to promote cancer cell quiescence by facilitating stabilization of p27. Purified quiescent B16 mouse melanoma cells expressed higher levels of miRNA-27b-3p and miRNA-455-3p relative to their proliferating counterparts. Induction of quiescence resulted in increased levels of these miRNAs in diverse types of human cancer cell lines. Inhibition of miRNA-27b-3p or miRNA-455-3p reduced, whereas its overexpression increased, the proportion of quiescent cells in the population, indicating that these miRNAs promote cancer cell quiescence. Accordingly, cancer xenografts bearing miRNA-27b-3p or miRNA-455-3p mimics were retarded in growth. miRNA-27b-3p targeted cyclin-dependent kinase regulatory subunit 1 (CKS1B), leading to reduction in p27 polyubiquitination mediated by S-phase kinase-associated protein 2 (Skp2).miRNA-455-3p targeted CDK2-associated cullin domain 1 (CAC1), which enhanced CDK2-mediated phosphorylation of p27 necessary for its polyubiquitination. Of note, the gene encoding miRNA-27b-3p was embedded in the intron of the chromosome 9 open reading frame 3 gene that was transcriptionally activated by p53. Similarly, the host gene of miRNA-455-3p, collagen alpha-1 (XXVII) chain, was also a p53 transcriptional target. Collectively, our results identify miRNA-27b-3p and miRNA-455-3p as important regulators of cancer cell quiescence in response to p53 and suggest that manipulating miRNA-27b-3p and miRNA-455-3p may constitute novel therapeutic avenues for improving outcomes of cancer treatment.Significance: Two novel p53-responsive microRNAs whose distinct mechanisms of action both stabilize p27 to promote cell quiescence and may serve as therapeutic avenues for improving outcomes of cancer treatment.

show abstract

Section: Cell Culturementioning

confidence: 99%

“…Western blotting and IP were carried out as described in previous publication (14). The relative intensity of bands was quantified using Image J and normalized against GAPDH.…”

Section: Western Blotting and Immunoprecipitationmentioning

confidence: 99%

A p53-Responsive miRNA Network Promotes Cancer Cell Quiescence

Liu

Farrelly

et al. 2018

Cancer Research

Self Cite

View full text Add to dashboard Cite

show abstract

“…Unlike normal cells, tumours produce high levels of oxidants due to chronically hyperactivated mitogenic and pro-survival signalling, as well as metabolic alterations, and have a greater reliance on MTH1 function. Thus, hMTH1 may play a crucial role in the survival of many cancers, such as melanoma and breast cancer (3,14), and the expression level of hMTH1 is positively correlated with the malignant degree of lung cancer and gastric carcinoma (15,16). Data were published in 2014 lending further support to the hypothesis that MTH1 has a role in the prevention of DNA damage and cancer cell survival.…”

Section: Introductionmentioning

confidence: 92%

Biological characterisation and application of human MTH1 and monoclonal antibody preparation

Chen

Li²,

et al. 2018

Oncol Rep

View full text Add to dashboard Cite

Human MutT homolog 1 (MTH1) hydrolyses oxidised nucleotide triphosphates, thereby preventing them from being incorporated into DNA; MTH1 has been found to be elevated in many types of cancers, including lung, stomach cancer, melanoma and breast cancer. Thus, tumour-targeted hMTH1 may be valuable for developing novel anticancer therapies. In the present study, we prepared human MTH1 protein and its monoclonal antibody (mAb). The hMTH1 gene was cloned into the prokaryotic expression vector pET28a and optimally expressed in the E. coli Transetta (DE3) strain. Using an Ni-NTA column and a G-50 gel filtration column, 20.1 mg of active hMTH1 was obtained from 1,000 ml of bacterial culture, and the purity was over 98%, as detected by high-performance liquid chromatography (HPLC). The half maximal inhibitory concentration (IC 50 ) of TH287 (hMTH1 inhibitor) was determined to be 3.53±0.47 nM using the recombinant hMTH1 protein (rhMTH1). The enzyme activity assay showed the Michaelis constant (K m ) and the catalytic constant (k cat ) of the protein were 106.13±48.83 µM and 3.64±0.58 sec -1 , respectively. The anti-hMTH1 mAb was obtained via the hybridoma technique and validated by western blot analysis. In addition, an immunofluorescence assay (IFA) and ELISA determined that the mAb could efficiently bind to natural hMTH1 expressed on the human breast cancer cell line MCF-7. Taken together, the results showed the rhMTH1 is an active protein and has practical applications for inhibitor selection, and our prepared hMTH1 mAb will provide a valuable tool for the further characterisation of hMTH1 and antitumour medicinal development in future.

show abstract

“…Human melanoma cell lines Me1007, Mel-RM, A375, Mel-CV, SK-Mel-28, and ME4405 were obtained as previously described (Jiang et al, 2008;Wang JY et al, 2017). They were cultured in DMEM containing 10% fetal bovine serum (lot FSP500; Excell Biology, Shanghai, China) and 0.1% Savelt (1:1,000; Hanheng Biotechnology, Shanghai, China).…”

Section: Cell Culture and Reagentsmentioning

confidence: 99%

Regulation of Cancer Stem Cell Self-Renewal by HOXB9 Antagonizes Endoplasmic Reticulum Stress-Induced Melanoma Cell Apoptosis via the miR-765–FOXA2 Axis

Lin

Zhang

Fan

et al. 2018

Journal of Investigative Dermatology

View full text Add to dashboard Cite

Adaptation to endoplasmic reticulum (ER) stress has been indicated as a driver of malignancy and resistance to therapy in human melanoma. However, the relationship between cancer stem cells and adaptation to ER stress remains unclear. Here, we show that the ratio of cancer stem cells is increased in ER stress-resistant melanoma cells, which inhibit ER stress-induced apoptosis and promote tumorigenesis. Further mechanistic studies showed that HOXB9 triggered by ER stress favors cancer stem cell self-renewal and enhances ER stress resistance. HOXB9 directly binds to the promoter of microRNA-765 and facilitates its transcription, which in turn targets FOXA2, resulting in a FOXA2 decrease and cancer stem cell increase. Additionally, an increase in HOXB9 promotes melanoma growth and inhibits cell apoptosis in a mouse xenograft model. Elevated HOXB9 is found in human melanoma tissues, which is associated with microRNA-765 up-regulation and FOXA2 decreases. Thus, our data showed that the HOXB9-dependent, microRNA-765-mediated FOXA2 pathway contributes to the survival of melanoma under ER stress by maintaining the properties of cancer stem cells.

show abstract

Skp2-Mediated Stabilization of MTH1 Promotes Survival of Melanoma Cells upon Oxidative Stress

Cited by 44 publications

References 48 publications

A p53-Responsive miRNA Network Promotes Cancer Cell Quiescence

A p53-Responsive miRNA Network Promotes Cancer Cell Quiescence

Biological characterisation and application of human MTH1 and monoclonal antibody preparation

Regulation of Cancer Stem Cell Self-Renewal by HOXB9 Antagonizes Endoplasmic Reticulum Stress-Induced Melanoma Cell Apoptosis via the miR-765–FOXA2 Axis

Contact Info

Product

Resources

About