Journal: Journal of Translational Medicine

Article Title: Exploring the oncogenic potential of circSOD2 in clear cell renal cell carcinoma: a novel positive feedback loop

doi: 10.1186/s12967-024-05290-9

Figure Lengend Snippet: miR-532-3p plays an inhibitory role in ccRCC by targeting PAX5. A The expression of miR-532-3p in RCC cells (769P, 786-O, A498, ACHN, Caki-1) and human renal cortical proximal convoluted tubule epithelial cells (HK2). B CCK8 assays demonstrated that cell proliferation was inhibited after transfected with miR-532-3p mimics. C Transwell assays demonstrated that cell migratory and invasive capacity was inhibited after transfected with miR-532-3p mimics. D FACS assays demonstrated that cell apoptosis was enhanced after transfected with miR-532-3p mimics. E Luciferase reporter assay in HEK293T cells co-transfected mimics miR- miR-532-3p or mimics NC and candidate genes of Luc-wild-type or Luc-mutant. F Western blot analysis indicated that miR-532-3p could down-regulate PAX5 expression in RCC cells. G Binding site of miR-532-3p and PAX5. (NS, nonsignificant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001)

Article Snippet: For transient transfection, RiboBio (Guangzhou, China) synthesized small interfering RNA (siRNA) targeting PAX5 along with a scrambled control.

Techniques: Expressing, Transfection, Luciferase, Reporter Assay, Mutagenesis, Western Blot, Binding Assay

Journal: Journal of Translational Medicine

Article Title: Exploring the oncogenic potential of circSOD2 in clear cell renal cell carcinoma: a novel positive feedback loop

doi: 10.1186/s12967-024-05290-9

Figure Lengend Snippet: Down-regulation of PAX5 suppresses proliferation, migration and invasion of RCC cells in vitro. A CCK8 assays demonstrated that cell proliferation was inhibited after PAX5 knockdown. B Transwell assays demonstrated that cell migratory and invasive capacity was inhibited after PAX5 knockdown. C FACS assays demonstrated that cell apoptosis was enhanced after PAX5 knockdown. D GSEA analysis in TCGA patients revealed that PAX5 may be involved in PI3K signaling. E Western blot analysis indicated that circSOD2 promotes the ccRCC progression through PI3K-AKT-mTOR pathway. ( A–C si-NC, referred to a scramble control of siRNA; si-1 & si-2, referred to siRNA targeting to PAX5 with different sequences; E sh-NC, referred to a scramble control of shRNA; sh-1 & sh-2, referred to shRNA targeting to circSOD2 with different sequences). (NS, nonsignificant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.)

Article Snippet: For transient transfection, RiboBio (Guangzhou, China) synthesized small interfering RNA (siRNA) targeting PAX5 along with a scrambled control.

Techniques: Migration, In Vitro, Knockdown, Western Blot, Control, shRNA

Journal: Journal of Translational Medicine

Article Title: Exploring the oncogenic potential of circSOD2 in clear cell renal cell carcinoma: a novel positive feedback loop

doi: 10.1186/s12967-024-05290-9

Figure Lengend Snippet: circSOD2 rescues the tumor suppressive effect of miR-532-3p. A Cell proliferation ability of 786-O transfected with control vector, miR-532-3p mimics alone or circSOD2 overexpressed plus miR-532-3p mimics. B Cell migration and invasion abilities of 786-O transfected with control vector, miR-532-3p mimics alone or circSOD2 overexpressed plus miR-532-3p mimics. C Cell proliferation ability of 769p transfected with control vector, miR-532-3p inhibitor alone or circSOD2 shRNA plus miR-532-3p inhibitor. D Cell migration and invasion abilities of 769p transfected with control vector, miR-532-3p inhibitor alone or circSOD2 shRNA plus miR-532-3p inhibitor. E Apoptosis of 786-O transfected with control vector, miR-532-3p mimics alone or circSOD2 overexpressed plus miR-532-3p mimics (above), and apoptosis of 769p transfected with control vector, miR-532-3p inhibitor alone or circSOD2 shRNA plus miR-532-3p inhibitor (below). F Western blot of PAX5 and p-PI3K levels after 769p transfected with control vector, miR-532-3p inhibitor alone or circSOD2 shRNA plus miR-532-3p inhibitor (left), and 786-O transfected with control vector, miR-532-3p mimics alone or circSOD2 overexpressed plus miR-532-3p mimics (right). G The diagram of the signal pathway. (NS, nonsignificant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001)

Article Snippet: For transient transfection, RiboBio (Guangzhou, China) synthesized small interfering RNA (siRNA) targeting PAX5 along with a scrambled control.

Techniques: Transfection, Control, Plasmid Preparation, Migration, shRNA, Western Blot

Journal: Journal of Translational Medicine

Article Title: Exploring the oncogenic potential of circSOD2 in clear cell renal cell carcinoma: a novel positive feedback loop

doi: 10.1186/s12967-024-05290-9

Figure Lengend Snippet: PAX5 enhances the biosynthesis of circSOD2 in ccRCC cells. A Venn diagram shows the intersection of transcription factors predicted by the three databases. B PAX5 has two possible transcription factor binding sites in the promoter of circSOD2. C qRT-PCR showed that the expression of circSOD2 decreased with the knockdown of the expression of PAX5 in 786-O cells. D Schematic diagram of luciferase reporter vectors containing mutations at each of these two sites (S1-MUT-Luc and S2-MUT-Luc). E Luciferase assays proved the the expression of PAX5 significantly activated the site 1, rather than the site 2. F ChIP assay showed that PAX5 was enriched at site 1. (NS, nonsignificant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001)

Article Snippet: For transient transfection, RiboBio (Guangzhou, China) synthesized small interfering RNA (siRNA) targeting PAX5 along with a scrambled control.

Techniques: Binding Assay, Quantitative RT-PCR, Expressing, Knockdown, Luciferase