Fig 1: CXCL8 blocks the apoptosis-promoting effects of miR-302c-3p and miR-520a-3p in CC cell lines. Flow cytometry shows the efficacy of pcDNA3.1-CXCL8 in restoring the proliferation-promoting and apoptosis-suppressive function in HeLa-S3 and C-33A cells. CXCL8 siRNA and pcDNA3.1-CXCL8 were compared with si-NC and pcDNA3.1, respectively. Cells co-transfected with miR-302c-3p mimic, miR-520a-3p mimic and pcDNA3.1-CXCL8 were compared with miR-302c-3p mimic and pcDNA3.1 as well as miR-520a-3p mimic and pcDNA3.1. *P<0.05; **P<0.01. CC, cervical carcinoma; miR, microRNA; si-NC, siRNA control; CXCL8, C-X-C motif ligand 8.
Fig 2: CXCL8 is a common target gene of miR-302c-3p and miR-520a-3p. (A) Expression levels of CXCL8 in tumour-adjacent and CC tissues were detected by reverse transcription-quantitative polymerase chain reaction. (B) Association between expression levels of CXCL8 and miR-302c-3p or miR-520a-3p was displayed with correlational analyses. (C) mRNA (top) and protein (bottom) expression levels of CXCL8 in CC cell lines (HeLa-S3 and C-33A cells) and normal cervical epithelial cells. The mRNA (D) and protein (E) expression level of CXCL8 in HeLa-S3 and C-33A cells after transfection with miR-302c-3p/miR-520a-3p mimics compared to blank cells. ‘miR-302c-3p mimic’ vs. ‘miR-302c-3p and miR-520a-3p’: P<0.05 (HeLa-S3), P<0.05 (C-33A); ‘miR-520a-3p mimic’ vs. ‘miR-302c-3p and miR-520a-3p’: P<0.05 (HeLa-S3), P<0.05 (C-33A), the values represent the gray values of the western blotting bands. (F) Diagram of the CXCL8 3'-UTR-containing reporter construct. Representative luciferase activity in 293T cells co-transfected with wild-type or mutated reporter plasmids and miR-302c-3p or miR-520a-3p mimics. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001. CC, cervical carcinoma; miR, microRNA; CXCL8, C-X-C motif ligand 8.
Fig 3: CXCL8 blocks the proliferation-suppressive effects of miR-302c-3p and miR-520a-3p in CC cell lines. (A) Cell counting kit-8 assay of HeLa-S3 and C-33A cells co-transfected with miR-302c-3p/miR-520a-3p mimics and/or CXCL8 plasmid. (B) EdU assays of HeLa-S3 (top) and C-33A (bottom) cells co-transfected with miR-302c-3p/miR-520a-3p mimics or CXCL8 plasmid. Graph shows the proportion of EdU positive cells. *P<0.05; **P<0.01; ***P<0.001. CC, cervical carcinoma; miR, microRNA; CXCL8, C-X-C motif ligand 8.
Fig 4: CXCL8 promotes proliferation and suppresses apoptosis in CC cells. (A) mRNA and protein expression levels of CXCL8 in HeLa-S3 cells and C-33A cells after transfection with si-NC and CXCL8 siRNA (si-CXCL8), pcDNA3.1 and pcDNA3.1-CXCL8, which were compared with si-NC group or pcDNA3.1 group, respectively. (B) Proliferation of HeLa-S3 and C-33A cells in response to transfection of CXCL8 siRNA or pcDNA3.1-CXCL8 or negative control plasmid. (C) EdU assay of HeLa-S3 and C-33A cells transfected with CXCL8 siRNA. Graph shows the proportion of EdU positive cells (EdU/DAPI). (D) Flow cytometry was used to determine the apoptosis of HeLa-S3 and C-33A cells transfected with CXCL8 siRNA. CXCL8 siRNA was compared with si-NC. *P<0.05; **P<0.01; ***P<0.001. CC, cervical carcinoma; CXCL8, C-X-C motif ligand 8; si-NC, siRNA control.
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