Research Paper Volume 13, Issue 3 pp 3945—3956

Figure 5. miR-205 targeted regulation of HOXD9. (A) Three major databases were used to predict target genes of miR-205. Based on the final biological function, HOXD9 was selected. (B) The correlation between miR-205 and HOXD9 mRNA expression was calculated in 100 pairs of TNBC specimens. (C) The correlation between miR-205 and HOXD9 protein expression was calculated in 100 pairs of TNBC specimens. (D) HOXD9 protein expression in miR-205 overexpression treated MDA-MB-231 cells was lower than that in normal MDA-MB-231 cells. (E) Western blot analysis of the HOXD9 protein and mRNA expression in miR-205 knockdown treated BT-549 cells and miR-205 overexpression treated MDA-MB-231 cells. (F) qRT-PCR analysis of the HOXD9 protein and mRNA expression in miR-205 knockdown treated BT-549 cells and miR-205 overexpression treated MDA-MB-231 cells. (G) Construction of HOXD9 mRNA 3’UTR wild-type and mutant plasmids using potential binding sites of miR-205 and HOXD9 mRNA. (H) Dual luciferase experiment was used to verify the binding of miR-205 and HOXD9 mRNA. (I) Western blot analysis of HOXD9 siRNA. (J) qRT-PCR analysis of HOXD9 siRNA. (K) Cell proliferation analysis of the miR-205 and HOXD9 knockdown treated BT-549 cells after different incubation time. (L) Cell proliferation analysis of the miR-205 and HOXD9 knockdown treated BT-549 cells with Cisplatin treatment. (M) Cell proliferation analysis of the miR-205 and HOXD9 knockdown treated BT-549 cells with Doxorubicin treatment. (N) Cell proliferation analysis of the miR-205 and HOXD9 knockdown treated BT-549 cells with Paclitaxel treatment. ***P<0.001.