Figure 1
From: CNN3 acts as a potential oncogene in cervical cancer by affecting RPLP1 mRNA expression
Highly expressed CNN3 promotes the viability and motility of cervical cancer cells. (A) Box plots of CNN3 mRNA expression in 20 cervical cancer tissues and 8 normal cervix tissues from Oncomine database of Pyeon Multi-cancer (*P < 0.05). (B) The mRNA expression of CNN3 in 7 pairs of cervical cancer and adjacent tissues by RT-PCR (*P < 0.05). Paired adjacent non-cancerous tissues were obtained at 3 cm away from the tumour edge and all tissues were reviewed by an experienced pathologist (Dr. Xiaofei Zhang). (C) CNN3 protein expression were then determined in 10 cell lines with immunoblot analysis. The grouping of blots cropped from same parts of the same gel. Full-length blots are presented in Supplementary Fig. S3. (D) SiHa and CaSki cells were transfected with si-NC, si-E5 and si-E6/E7 siRNA for 48 h. The expressions of P53 and CNN3 protein were detected by immunoblot analysis. The expressions of P53, a downstream molecule of 16E6, were tested to confirm the interference efficiency of si-E6/E727,28. Blots of CNN3 and β-actin cropped from same parts of the same gel, while blots of P53 cropped from different parts of gel. Full-length blots are presented in Supplementary Fig. S3. (E) Cell viability of SiHa and CaSki was detected by CCK8 assays after transfection with NC-plasmid, CNN3-plasmid, si-NC, si-CNN3#1 and si-CNN3#2 for 24 h. The OD value (450 nm) was detected every 24 h and continuously tested for 5 days. The results represent the mean ± SEM (n = 3). ***P < 0.001, ###P < 0.001. (F) Representative photographs of colony formation assays of SiHa and CaSki after transfection with NC-plasmid, CNN3-plasmid, si-NC, si-CNN3#1 and si-CNN3#2 are shown (upper panels), respectively. Quantification and statistics of colony area percentage per well is shown (lower graphs), mean ± SEM (n ≥ 3). *P < 0.05, **P < 0.01. (G) Boyden chamber migration and invasion assays were performed to detect the migration and invasiveness ability of SiHa and CaSki cells after transfection with NC-plasmid, CNN3-plasmid, si-NC, si-CNN3#1 and si-CNN3#2 for 24 h, respectively. Representative images (scale bars, 100 μm) and corresponding number (SiHa) or area (CaSki) of migrated (or invaded) cells from three independent experiments are shown, mean ± SEM (n ≥ 3), *P < 0.05, **P < 0.01, ***P < 0.001.
