Research Paper Volume 14, Issue 11 pp 4755—4768

LINC00094/miR-19a-3p/CYP19A1 axis affects the sensitivity of ER positive breast cancer cells to Letrozole through EMT pathway

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Figure 6. Adsorption of miR-19a-3p by LINC00094 sponge affects the expression of CYP19A1. (A) miRNA-specific PCR was used to detect the expression level of miR-19a-3p (U6 was an internal reference). ****P<0.0001. (B) mRNA expression of CYP19A1 in each transfection group of breast cancer cells was detected by RT-QPCR (β-actin as internal reference). ****P<0.0001. (C) Immunofluorescence analysis of DAPI and FITC-CYP19A1 expression in each transfection group of BT-474 cells, with the scale of 10 μm. (D) Western Blot was used to detect the protein expression level of CYP19A1 in each transfection group of MCF-7, with GAPDH as internal reference. The experiment was repeated for 3 times. ****P<0.0001. (E) Western Blot was used to detect the protein expression level of CYP19A1 in BT-474 transfected cells, with GAPDH as internal reference, and the experiment was repeated for 3 times. *P<0.05, ****P<0.0001. (F) The expression levels of CYP19A1 (β-actin as internal reference) and miR-19a-3p (U6 as internal reference) in plasma of breast cancer patients were detected by RT-QPCR. *P<0.05. (G) Binding sequence of miR-19a-3p to CYP19A1 3’UTR and mutation site of mutant CYP19A1 3’UTR plasmid. (H) Luciferase activity assay of HEK-293T cells in each transfection group. ***P<0.001.