Figure 4
hsa-miR-4756-3p regulated TNBC metastasis in vitro and in vivo through FOXM1-TGFβ1-Smad3-EMT pathway. (A) Control sgRNA and FOMX1 KO were transfected in MDA-MB-231 cells, after single clone selection, using western blot to detect change of FOXM1 expression. (B) MDA-MB-231 cells was divided into control, hsa-miR-4756-3p inhibitor, hsa-miR-4756-3p inhibitor + FOXM1 KO group, control group transfected with control miRNA, hsa-miR-4756-3p inhibitor transfected with hsa-miR-4756-3p inhibitor, hsa-miR-4756-3p inhibitor + FOXM1 KO group was 231-FOXM1 KO transfected with hsa-miR-4756-3p inhibitor, then using wound healing assay to assess the migration change. (C) Employing QPCR and western blot to find the hsa-miR-4756-3p (left) and FOXM1(right) expression in 231 and trained 231 cells. (D) 15 nude mice were divided into control, hsa-miR-4756-3p inhibitor, hsa-miR-4756-3p inhibitor + FOXM1 KO group, control, hsa-miR-4756-3p inhibitor group were injected trained 231 in mammary gland fat pad, hsa-miR-4756-3p inhibitor + FOXM1 KO group were injected with FOXM1 KO trained 231 cells, then control miRNA was injected control nude mice using DOPC liposomes, hsa-miR-4756-3p inhibitor was injected in other 2 groups, after 2 months, mice lung metastasis were detected. (E) In control, hsa-miR-4756-3p inhibitor, hsa-miR-4756-3p inhibitor + FOXM1 KO group nude mice primary tumor, protein was extracted and TGFβ1 signal pathway, EMT pathway were assess.
