MITF regulates the subcellular location of HIF1α through SUMOylation to promote the invasion and metastasis of daughter cells derived from polyploid giant cancer cells

Zheng,Minying; Tian,Shifeng; Zhou,Xinyue; Yan,Man; Zhou,Mingming; Yu,Yongjun; Zhang,Yue; Wang,Xiaorui; Li,Na; Ren,Li; Zhang,Shiwu

doi:10.3892/or.2024.8722

MITF regulates the subcellular location of HIF1α through SUMOylation to promote the invasion and metastasis of daughter cells derived from polyploid giant cancer cells

Authors:
- Minying Zheng
- Shifeng Tian
- Xinyue Zhou
- Man Yan
- Mingming Zhou
- Yongjun Yu
- Yue Zhang
- Xiaorui Wang
- Na Li
- Li Ren
- Shiwu Zhang
View Affiliations

Affiliations: Department of Pathology, Tianjin Union Medical Center, Tianjin 300121, P.R. China, Graduate School, Tianjin Medical University, Tianjin 300070, P.R. China, Graduate School, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, P.R. China, Department of Clinical Laboratory, Tianjin Medical University Cancer Institution and Hospital, Tianjin 300090, P.R. China
Published online on: March 6, 2024 https://doi.org/10.3892/or.2024.8722
Article Number: 63
Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

High concentrations of cobalt chloride (CoCl₂) can induce the formation of polyploid giant cancer cells (PGCCs) in various tumors, which can produce daughter cells with strong proliferative, migratory and invasive abilities via asymmetric division. To study the role of hypoxia‑inducible factor (HIF) 1α in the formation of PGCCs, colon cancer cell lines Hct116 and LoVo were used as experimental subjects. Western blotting, nuclear and cytoplasmic protein extraction and immunocytochemical experiments were used to compare the changes in the expression and subcellular localization of HIF1α, microphthalmia‑associated transcription factor (MITF), protein inhibitor of activated STAT protein 4 (PIAS4) and von Hippel‑Lindau disease tumor suppressor (VHL) after treatment with CoCl₂. The SUMOylation of HIFα was verified by co‑immunoprecipitation assay. After inhibiting HIF1α SUMOylation, the changes in proliferation, migration and invasion abilities of Hct116 and LoVo were compared by plate colony formation, wound healing and Transwell migration and invasion. In addition, lysine sites that led to SUMOylation of HIF1α were identified through site mutation experiments. The results showed that CoCl₂ can induce the formation of PGCCs with the expression level of HIF1α higher in treated cells than in control cells. HIF1α was primarily located in the cytoplasm of control cell. Following CoCl₂ treatment, the subcellular localization of HIF1α was primarily in the nuclei of PGCCs with daughter cells (PDCs). After treatment with SUMOylation inhibitors, the nuclear HIF1α expression in PDCs decreased. Furthermore, their proliferation, migration and invasion abilities also decreased. After inhibiting the expression of MITF, the expression of HIF1α decreased. MITF can regulate HIF1α SUMOylation. Expression and subcellular localization of VHL and HIF1α did not change following PIAS4 knockdown. SUMOylation of HIF1α occurs at the amino acid sites K391 and K477 in PDCs. After mutation of the two sites, nuclear expression of HIF1α in PDCs was reduced, along with a significant reduction in the proliferation, migration and invasion abilities. In conclusion, the post‑translation modification regulated the subcellular location of HIF1α and the nuclear expression of HIF1α promoted the proliferation, migration and invasion abilities of PDCs. MITF could regulate the transcription and protein levels of HIF1α and participate in the regulation of HIF1α SUMOylation.

View Figures

View References

1	Fan L, Zheng M, Zhou X, Yu Y, Ning Y, Fu W, Xu J and Zhang S: Molecular mechanism of vimentin nuclear localization associated with the migration and invasion of daughter cells derived from polyploid giant cancer cells. J Transl Med. 21:7192023. View Article : Google Scholar : PubMed/NCBI
2	Li Z, Zheng M, Zhang H, Yang X, Fan L, Fu F, Fu J, Niu R, Yan M and Zhang S: Arsenic trioxide promotes tumor progression by inducing the formation of PGCCs and embryonic hemoglobin in colon cancer cells. Front Oncol. 11:7208142021. View Article : Google Scholar : PubMed/NCBI
3	Zhang S, Mercado-Uribe I, Xing Z, Sun B, Kuang J and Liu J: Generation of cancer stem-like cells through the formation of polyploid giant cancer cells. Oncogene. 33:116–128. 2014. View Article : Google Scholar : PubMed/NCBI
4	Zhao Q, Zhang K, Li Z, Zhang H, Fu F, Fu J, Zheng M and Zhang S: High migration and invasion ability of PGCCs and their daughter cells associated with the nuclear localization of S100A10 modified by SUMOylation. Front Cell Dev Biol. 9:6968712021. View Article : Google Scholar : PubMed/NCBI
5	Nehme Z, Pasquereau S, Haidar Ahmad S, El Baba R and Herbein G: Polyploid giant cancer cells, EZH2 and Myc upregulation in mammary epithelial cells infected with high-risk human cytomegalovirus. EBioMedicine. 80:1040562022. View Article : Google Scholar : PubMed/NCBI
6	Lv H, Shi Y, Zhang L, Zhang D, Liu G, Yang Z, Li Y, Fei F and Zhang S: Polyploid giant cancer cells with budding and the expression of cyclin E, S-phase kinase-associated protein 2, stathmin associated with the grading and metastasis in serous ovarian tumor. BMC Cancer. 14:5762014. View Article : Google Scholar : PubMed/NCBI
7	Bowers RR, Andrade MF, Jones CM, White-Gilbertson S, Voelkel-Johnson C and Delaney JR: Autophagy modulating therapeutics inhibit ovarian cancer colony generation by polyploid giant cancer cells (PGCCs). BMC Cancer. 22:4102022. View Article : Google Scholar : PubMed/NCBI
8	Pustovalova M, Blokhina T, Alhaddad L, Chigasova A, Chuprov-Netochin R, Veviorskiy A, Filkov G, Osipov AN and Leonov S: CD44+ and CD133+ non-small cell lung cancer cells exhibit DNA damage response pathways and dormant polyploid giant cancer cell enrichment relating to their p53 status. Int J Mol Sci. 23:49222022. View Article : Google Scholar : PubMed/NCBI
9	Bilé-Silva A, Lopez-Beltran A, Rasteiro H, Vau N, Blanca A, Gomez E, Gaspar F and Cheng L: Pleomorphic giant cell carcinoma of the prostate: Clinicopathologic analysis and oncological outcomes. Virchows Arch. 482:493–505. 2023. View Article : Google Scholar : PubMed/NCBI
10	Wicks EE and Semenza GL: Hypoxia-inducible factors: Cancer progression and clinical translation. J Clin Invest. 132:e1598392022. View Article : Google Scholar : PubMed/NCBI
11	Konopleva MY and Jordan CT: Leukemia stem cells and microenvironment: Biology and therapeutic targeting. J Clin Oncol. 29:591–599. 2011. View Article : Google Scholar : PubMed/NCBI
12	Paredes F, Williams HC and San Martin A: Metabolic adaptation in hypoxia and cancer. Cancer Lett. 502:133–142. 2021. View Article : Google Scholar : PubMed/NCBI
13	Zhao H, Wang X and Fang B: HIF1A promotes miR-210/miR-424 transcription to modulate the angiogenesis in HUVECs and HDMECs via sFLT1 under hypoxic stress. Mol Cell Biochem. 477:2107–2119. 2022. View Article : Google Scholar : PubMed/NCBI
14	Zhang S, Mercado-Uribe I, Hanash S and Liu J: iTRAQ-based proteomic analysis of polyploid giant cancer cells and budding progeny cells reveals several distinct pathways for ovarian cancer development. PLoS One. 8:e801202013. View Article : Google Scholar : PubMed/NCBI
15	Semenza GL: Targeting HIF-1 for cancer therapy. Nat Rev Cancer. 3:721–732. 2003. View Article : Google Scholar : PubMed/NCBI
16	Wei J, Yang Y, Lu M, Lei Y, Xu L, Jiang Z, Xu X, Guo X, Zhang X, Sun H and You Q: Recent advances in the discovery of HIF-1α-p300/CBP inhibitors as anti-cancer agents. Mini Rev Med Chem. 18:296–309. 2018. View Article : Google Scholar : PubMed/NCBI
17	Du L, Liu W and Rosen ST: Targeting SUMOylation in cancer. Curr Opin Oncol. 33:520–525. 2021. View Article : Google Scholar : PubMed/NCBI
18	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
19	Itokawa H, Totsuka N, Nakahara K, Takeya K, Lepoittevin JP and Asakawa Y: Antitumor principles from ginkgo biloba L. Chem Pharm Bull (Tokyo). 35:3016–3020. 1987. View Article : Google Scholar : PubMed/NCBI
20	Liu K, Wang X, Li D, Xu D, Li D, Lv Z, Zhao D, Chu WF and Wang XF: Ginkgolic acid, a SUMO-1 inhibitor, inhibits the progression of oral squamous cell carcinoma by alleviating SUMOylation of SMAD4. Mol Ther Oncolytics. 16:86–99. 2020. View Article : Google Scholar : PubMed/NCBI
21	Buscà R, Berra E, Gaggioli C, Khaled M, Bille K, Marchetti B, Thyss R, Fitsialos G, Larribère L, Bertolotto C, et al: Hypoxia-inducible factor 1{alpha} is a new target of microphthalmia-associated transcription factor (MITF) in melanoma cells. J Cell Biol. 170:49–59. 2005. View Article : Google Scholar : PubMed/NCBI
22	Cai Q, Verma SC, Kumar P, Ma M and Robertson ES: Hypoxia inactivates the VHL tumor suppressor through PIASy-mediated SUMO modification. PLoS One. 5:e97202010. View Article : Google Scholar : PubMed/NCBI
23	Bae SH, Jeong JW, Park JA, Kim SH, Bae MK, Choi SJ and Kim KW: Sumoylation increases HIF-1alpha stability and its transcriptional activity. Biochem Biophys Res Commun. 324:394–400. 2004. View Article : Google Scholar : PubMed/NCBI
24	Haidar Ahmad S, El Baba R and Herbein G: Polyploid giant cancer cells, cytokines and cytomegalovirus in breast cancer progression. Cancer Cell Int. 23:1192023. View Article : Google Scholar : PubMed/NCBI
25	Casotti MC, Meira DD, Zetum ASS, Araújo BC, Silva DRCD, Santos EVWD, Garcia FM, Paula F, Santana GM, Louro LS, et al: Computational biology helps understand how polyploid giant cancer cells drive tumor success. Genes (Basel). 14:8012023. View Article : Google Scholar : PubMed/NCBI
26	Alhaddad L, Chuprov-Netochin R, Pustovalova M, Osipov AN and Leonov S: Polyploid/multinucleated giant and slow-cycling cancer cell enrichment in response to X-ray irradiation of human glioblastoma multiforme cells differing in radioresistance and TP53/PTEN status. Int J Mol Sci. 24:12282023. View Article : Google Scholar : PubMed/NCBI
27	Hoellein A, Fallahi M, Schoeffmann S, Steidle S, Schaub FX, Rudelius M, Laitinen I, Nilsson L, Goga A, Peschel C, et al: Myc-induced SUMOylation is a therapeutic vulnerability for B-cell lymphoma. Blood. 124:2081–2090. 2014. View Article : Google Scholar : PubMed/NCBI
28	Driscoll JJ, Pelluru D, Lefkimmiatis K, Fulciniti M, Prabhala RH, Greipp PR, Barlogie B, Tai YT, Anderson KC, Shaughnessy JD Jr, et al: The sumoylation pathway is dysregulated in multiple myeloma and is associated with adverse patient outcome. Blood. 115:2827–2834. 2010. View Article : Google Scholar : PubMed/NCBI
29	Xia QD, Sun JX, Xun Y, Xiao J, Liu CQ, Xu JZ, An Y, Xu MY, Liu Z, Wang SG and Hu J: SUMOylation pattern predicts prognosis and indicates tumor microenvironment infiltration characterization in bladder cancer. Front Immunol. 13:8641562022. View Article : Google Scholar : PubMed/NCBI
30	Dai X, Zhang T and Hua D: Ubiquitination and SUMOylation: Protein homeostasis control over cancer. Epigenomics. 14:43–58. 2022. View Article : Google Scholar : PubMed/NCBI
31	Bettermann K, Benesch M, Weis S and Haybaeck J: SUMOylation in carcinogenesis. Cancer Lett. 316:113–125. 2012. View Article : Google Scholar : PubMed/NCBI
32	Han ZJ, Feng YH, Gu BH, Li YM and Chen H: The post-translational modification, SUMOylation, and cancer (review). Int J Oncol. 52:1081–1094. 2018.PubMed/NCBI
33	Levy C, Khaled M and Fisher DE: MITF: Master regulator of melanocyte development and melanoma oncogene. Trends Mol Med. 12:406–414. 2006. View Article : Google Scholar : PubMed/NCBI
34	Nooron N, Ohba K, Takeda K, Shibahara S and Chiabchalard A: Dysregulated expression of MITF in subsets of hepatocellular carcinoma and cholangiocarcinoma. Tohoku J Exp Med. 242:291–302. 2017. View Article : Google Scholar : PubMed/NCBI
35	Li W, Qin X, Wang B, Xu G, Zhang J, Jiang X, Chen C, Qiu F and Zou Z: MiTF is associated with chemoresistance to cisplatin in A549 lung cancer cells via modulating lysosomal biogenesis and autophagy. Cancer Manag Res. 12:6563–6573. 2020. View Article : Google Scholar : PubMed/NCBI
36	Wilkinson KA and Henley JM: Mechanisms, regulation and consequences of protein SUMOylation. Biochem J. 428:133–145. 2010. View Article : Google Scholar : PubMed/NCBI
37	Sun Y, Perera J, Rubin BP and Huang J: SYT-SSX1 (synovial sarcoma translocated) regulates PIASy ligase activity to cause overexpression of NCOA3 protein. J Biol Chem. 286:18623–18632. 2011. View Article : Google Scholar : PubMed/NCBI
38	Mabb AM, Wuerzberger-Davis SM and Miyamoto S: PIASy mediates NEMO sumoylation and NF-kappaB activation in response to genotoxic stress. Nat Cell Biol. 8:986–993. 2006. View Article : Google Scholar : PubMed/NCBI
39	Chien W, Lee KL, Ding LW, Wuensche P, Kato H, Doan NB, Poellinger L, Said JW and Koeffler HP: PIAS4 is an activator of hypoxia signalling via VHL suppression during growth of pancreatic cancer cells. Br J Cancer. 109:1795–1804. 2013. View Article : Google Scholar : PubMed/NCBI