Fig 1: high expression levels of miR-543 predicts the poor prognosis of HNSC patients. (A) Data from TCGA demonstrated that when compared with normal tissues, CYP3A5 exhibited lower expression levels in cancerous tissues. (B) Low expression levels of CYP3A5 in the individual cancer stages. (C) Low expression levels of CYP3A5 in the tumor grades. (D) High expression levels of miR-543 predicted poor prognosis in HNSC patients. Data from TCGA revealed that miR-543 at a high level of expression equated to a poorer survival prognosis in HSNC patients. TCGA, The Cancer Genome Atlas; miR, microRNA; CYP3A5, cytochrome P450 family 3 subfamily A member 5; HNSC, head and neck squamous cell carcinoma.
Fig 2: CYP3A5 is negatively associated with miR-543 expression in OSCC. (A and B) RT-qPCR analysis of the level of CYP3A5 revealed a decrease in cancerous tissues when compared with that in the adjacent non-tumor tissues according to the results detected in 20 paired clinical samples. (C) Analysis of the correlation of expression levels of CYP3A5 and miR-543 in 20 pairs of OSCC. The results revealed that there was a significant negative correlation between CYP3A5 and miR-543. (D) RT-qPCR analysis determined that CYP3A5 was significantly decreased in the 3 OSCC cell lines when compared with HOK cells. **P<0.01, ***P<0.001. miR, microRNA; OSCC, oral squamous cell carcinoma; CYP3A5, cytochrome P450 family 3 subfamily A member 5; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
Fig 3: CYP3A5 is a direct target gene of miR-543. (A) The screened target genes were initially validated in OSCC cell lines, and it was determined that the expression level of CYP3A5 had an inverse correlation with miR-543 in the OSCC cell lines. (B) The sequence of the miR-543 binding site within the 3'UTR of human CYP3A5. (C) CYP3A5 had a direct binding role with miR-543. Analysis of the relative luciferase activities of CYP3A5-WT and CYP3A5-MUT in 293T cells. The results revealed that overexpression of miR-543 significantly reduced the luciferase activity of WT CYP3A5 3'UTR, while the MUT CYP3A5 3'UTR had virtually no change. CYP3A5 (D) mRNA and (F) protein expression were decreased in SCC9, SCC25 and CAL27 cells following treatment with miR-543 mimic when compared with the mimic NC. CYP3A5 (E) mRNA and (G) protein expression were decreased in SCC9, SCC25 and CAL27 cells following treatment with the miR-543 inhibitor when compared with the inhibitor NC. *P<0.05, **P<0.01, ***P<0.001; ns, not significant; miR, microRNA; OSCC, oral squamous cell carcinoma; NC, negative control; CYP3A5, cytochrome P450 family 3 subfamily A member 5; UTR, untranslated region; MUT, mutant; WT, wild-type.
Fig 4: Immunohistochemical analysis of the expression levels of CYP3A5; the expression was low in OSCC tissues when compared with that in the adjacent non-tumor tissues. *P<0.05, **P<0.01. CYP3A5, cytochrome P450 family 3 subfamily A member 5.
Fig 5: Effects of CYP3A5 on the phenotypes of SCC9 cells. (A) CYP3A5 expression was significantly downregulated in SCC9 cells following transfection with CYP3A5 siRNA when compared with NC. Knockdown of CYP3A5 in SCC9 cells promoted (B) cell proliferation and (C) invasion. *P<0.05, **P<0.01; ns, not significant; NC, negative control; CYP3A5, cytochrome P450 family 3 subfamily A member 5; siRNA, small interfering.
Supplier Page from Abcam for Anti-CYP3A5 antibody [EPR4396]