https://orcid.org/0000-0002-1439-582X
Figures & data
Table 1. Primers for RNF220 and reference genes
Figure 1. RNF220 was highly expressed in colorectal cancer tissues and cells. (a) TCGA database showed the expression level of RNF220 in primary colorectal tumor (n = 286) and normal tissues (n = 41), p < 0.001. (b-c) RT-PCR and western blot analysis showed the RNF220 mRNA and protein level in colorectal tumor and para-carcinoma tissue from 65 patients, p < 0.001, β-actin was normalized as an internal control. (d-e) RT-PCR and western blot analysis showed the RNF220 protein level in normal colonic mucosa NCM460 cells and colorectal cancer cells SW480, HCT116, SW620 and SW837, p < 0.05, p < 0.01, p < 0.001. All representative data are from three independent experiments. β-actin was normalized as an internal control
Figure 2. RNF220 promoted the proliferation, migration and invasion of colorectal cancer cells. (a) Western blot assay detected the transfection efficiency, p < 0.01, p < 0.001. β-actin was normalized as an internal control. (b-c) Brdu and colon formation assays dectected the SW480 cells proliferasion abilities in sh-NC, sh-RNF220, pcDNA and pcDNA-RNF220 groups. p < 0.05, p < 0.01. (d-e) Transwell and wound healing assays examined the SW480 cells invasion and migration in sh-NC, sh-RNF220, pcDNA and pcDNA-RNF220 groups. p < 0.05, p < 0.01, p < 0.001. All representative data are from three independent experiments
Figure 3. RNF220 promotes the stemness of colorectal cancer cells. (a) Representative images of sphere formation induced by the transfection of sh-NC, sh-RNF220, pcDNA and pcDNA-RNF220 plasmids into SW480 cells. (b) Western blot analysis shows the expression levels of SOX2, OCT4 and NANOG in sh-NC, sh-RNF220, pcDNA and pcDNA-RNF220 groups, p < 0.05, p < 0.01, p < 0.001. All representative data are from three independent experiments. β-actin was normalized as an internal control
Figure 4. RNF220 regulates BMI1 expression through USP22. (a) Western blot analysis examined the USP22 and BMI1 expression in SW480 cells transfected with sh-NC, sh-RNF220, pcDNA and pcDNA-RNF220 plasmids, p < 0.01, p < 0.001. β-actin was normalized as an internal control. (b) Western blot analysis examined the USP22 and BMI1 expression in SW480 cells transfected with sh-NC+pcDNA, sh- RNF220+ pcDNA and sh-RNF220+ pcDNA-USP22 plasmids, p < 0.01, p < 0.001. All representative data are from three independent experiments. β-actin was normalized as an internal control
Figure 5. RNF220 promotes tumor growth in vivo. (a) Tumors were obtatined from sh-ctrl and shRNA-RNF220 groups of subcutaneous xenograft mice model (n = 5) and tumors volume and weight were measured after injection for 30 days, p < 0.001. (b) The expression of RNF220, USP22, BMI1 and PCNA in shRNA-RNF220 group mice and sh-ctrl group mice were detected by western blot, p < 0.001. All representative data are from three independent experiments. β-actin was normalized as an internal control
Availability of data and materials
All data generated or analyzed during this study are included in this published article.