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. 2024 Mar 6;51(5):63. doi: 10.3892/or.2024.8722

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Copyright: © 2024 Zheng et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 4. — MITF regulates the SUMOylation of HIF1α. (A) The interaction between HIF1α and MITF was verified by Co-IP in (a) Hct116 and (b) LoVo control cells and PDCs. (B) mRNA expression levels of (a and c) MITF and (b and d) HIF1α were detected in Hct116 and LoVo PDCs following MITF knockdown. (C) (a) Hct116 and (b) LoVo PDCs were treated with ML at different concentrations (40, 80, 160 and 320 µM). At each time point (0, 20, 40, 60 and 80 h), cell viability was assessed using CCK8 assay. (D) Western blotting showing the (a) total, (b) cytoplasmic and (c) nuclear expression of MITF, HIF1α, SUMO1 and SUMO2/3 in Hct116 and LoVo PDCs following treatment with different concentrations of ML. (d) Western blotting showing the expression of VHL in Hct116 and LoVo PDCs following treatment with differernt concentrations of ML. (E) Colony formation of 50, 100 and 150 (a) Hct116 and (b) LoVo PDCs before and after ML treatment. (F) Wound healing assay of (a) Hct116 and (b) LoVo PDCs before and after ML treatment at 0 h and 24 h (magnification, ×100). (G) (a) The differences in colony formation efficiency of Hct116 and LoVo PDCs before and after ML treatment. (b) Statistical analysis of wound healing index of Hct116 and LoVo PDCs before and after ML treatment. (H) The invasion and migration abilities of (a) Hct116 and (b) LoVo PDCs before and after ML treatment. (I) Comparison of the average cell number in invasion and migration assay of (a) Hct116 and (b) LoVo PDCs before and after ML treatment. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. HIF1α, hypoxia inducible factor 1 alpha; MITF, microphthalmia-associated transcription factor; SUMO, small ubiquitin-like modifier; Co-IP, co-immunoprecipitation; PDCs, daughter cells; ML, methyl linoleate; PGCCs, polyploid giant cells; Tre, PGCCs with PDCs; Ctr, control.