Fig 1: DCUN1D1 is a target gene of miR-195 in cervical cancer cells. (A) DCUN1D1 was predicted to be a putative target gene of miR-195. (b) The WT and MuT DCUN1D1 3'-UTR luciferase reporter plasmids were generated. (C) Luciferase reporter gene assay data indicated that the luciferase activity was significantly reduced in the presence of miR-195 in HeLa and SiHa cells transfected with WT DCUN1D1 3'-UTR luciferase reporter plasmid, but not with MuT DCUN1D1 3'-UTR luciferase reporter plasmid. **P<0.01 vs. control. (D) Reverse transcription-quantitative polymerase chain reaction and (E) western blotting were used to examine the mRNA and protein expression levels of DCUN1D1 in HeLa and SiHa cells transfected with miR-195 mimic or miR-NC, respectively. **P<0.01 vs. miR-NC. The experiments were repeated three times. DCUN1D1, defective in cullin neddylation 1 domain containing 1; miR, microRNA; WT, wild-type; MuT, mutant type; miR-NC, negative control miR mimic; CDS, coding sequence; UTR, untranslated region.
Fig 2: Upregulation of DCUN1D1 is associated with aggressive progression and poor prognosis in cervical cancer. (A) Reverse transcription-quantitative polymerase chain reaction was used to examine the mRNA expression levels of DCUN1D1 in 72 cervical cancer tissue samples compared with their matched adjacent non-tumor tissue samples. ****P<0.0001 vs. adjacent. (b) A significant negative correlation was demonstrated between the expression levels of miR-195 and DCUN1D1 in these 72 cervical cancer tissue samples. (C) The cervical cancer patients with high expression levels of DCUN1D1 exhibited shorter survival times compared with those with low expression levels of DCUN1D1. The experiments were repeated three times. DCUN1D1, defective in cullin neddylation 1 domain containing 1; miR, microRNA; mRNA, messenger RNA.
Fig 3: HPV16 E6 regulates the expression levels of miR-195 and DCUN1D1 in cervical cancer cells. (A) Reverse transcription-quantitative polymerase chain reaction was used to examine the miR-195 expression levels in (A) cervical cancer cell lines, including HeLa (HPV18+), SiHa (HPV16+) and C33A (HPV-), and (b) HeLa and SiHa cells transfected with HPV16 E6 siRNA, HPV16 E7 siRNA, HPV18 E6 siRNA and HPV18 E7 siRNA, respectively. **P<0.01 vs. C33A. (C) Western blot analysis was performed to examine the protein expression level of DCUN1D1 in SiHa and HeLa cells transfected with HPV16 E6 siRNA or HPV18 E6 siRNA, respectively. The experiments were repeated three times. HPV, human papilloma virus; miR, microRNA; DCUN1D1, defective in cullin neddylation 1 domain containing 1; siRNA, small interfering RNA; NC, negative control.
Fig 4: DCUN1D1 restored miR-195-mediated cell migration and invasion in cervical cancer. The miR-195-overexpressing HeLa and SiHa cells were transfected with pcDNA3.1-DCUN1D1 expression plasmid, or with blank pcDNA3.1 vector as the control group. Following transfection, (A) wound healing (magnification, ×40) and (b) Transwell (magnification, ×400) assays were used to examine cell migration and invasion, respectively. **P<0.01 vs. miR-195 + blank. The experiments were repeated three times. DCUN1D1, defective in cullin neddylation 1 domain containing 1; miR, microRNA; mRNA, messenger RNA.
Fig 5: DCUN1D1 restored miR-195-mediated cell proliferation in cervical cancer. The miR-195-overexpressing HeLa and SiHa cells were transfected with pcDNA3.1-DCUN1D1 expression plasmid, or with blank pcDNA3.1 vector as the control group. Following transfection, (A) reverse transcription-quantitative polymerase chain reaction and (B) western blot analysis were performed to examine the mRNA and protein expression levels of DCUN1D1. (C) MTT assay was used to examine cell proliferation. **P<0.01 vs. miR-195 + blank. The experiments were repeated three times. DCUN1D1, defective in cullin neddylation 1 domain containing 1; miR, microRNA; mRNA, messenger RNA.
Supplier Page from Abcam for Anti-DCUN1D1 antibody [EPR13492]