Research Paper Volume 16, Issue 6 pp 5526—5544

Cinobufotalin regulates the USP36/c-Myc axis to suppress malignant phenotypes of colon cancer cells in vitro and in vivo

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Figure 6. Suppressive effect of CBF on malignant phenotypes of colon cancer cells is mediated by USP36. (A) The mRNA (top) and protein (button) expression levels of USP36 in HCT116 cells were respectively detected by RT-PCR and western blot to validate the transfection efficiency of sh-USP36. (B, C) The cell proliferation was detected by colony formation (B) and Edu staining (C, Scale bar = 100 μm) assays. (D) The cell viability was detected by CCK-8 assay after a series of concentrations of Oxa for the determination of IC50 value to Oxa. (E) The cellular self-renewal capacity was investigated by sphere formation assay (Scale bar=100 μm). (F) The protein expression of cancer stem cell-related markers (CD133, CD44, Nanog, and Oct-4) was detected by western blot. (G) The cell migration was detected by wound healing assay (Scale bar = 100 μm). (H) The cell invasion was observed by Transwell assay (Scale bar = 100 μm). *p < 0.05 and **p < 0.01; Abbreviation: ns: no significance.