Research Paper Volume 13, Issue 7 pp 10431—10449

miR-224 aggravates cancer-associated fibroblast-induced progression of non-small cell lung cancer by modulating a positive loop of the SIRT3/AMPK/mTOR/HIF-1α axis

Effects of miR-224 in CAFs-induced promotive effect on NSCLC cells. (A, B) CAFs were transfected with miR-224 mimics or inhibitors for 24 hours, and the level of miR-224 in CAFs (A) and the co-culture medium of CAFs-NSCLCs (B) was detected by RT-PCR. (C) The level of miR-224 in NSCLC cells co-culture with CAFs for 24 hours were detected by RT-PCR. (D) The proliferation of NSCLC cells co-culture with CAFs for 24 hours was determined by the CCK8 assay. (E) Colony formation assay was used to evaluate the cell colony of NSCLC cells co-culture with CAFs for 24 hours. (F) Transwell assay was used to test the invasive ability of NSCLC cells co-culture with CAFs for 24 hours. (G) EMT markers including E-cadherin, Vimentin, and N-cadherin of NSCLC cells co-culture with CAFs for 24 hours were determined by western blot. (H) Tube formation assay was conducted to monitor the tube formation ability of HUVECs co-culture with CAFs for 24 hours. *, ** and *** represents P

Figure 3. Effects of miR-224 in CAFs-induced promotive effect on NSCLC cells. (A, B) CAFs were transfected with miR-224 mimics or inhibitors for 24 hours, and the level of miR-224 in CAFs (A) and the co-culture medium of CAFs-NSCLCs (B) was detected by RT-PCR. (C) The level of miR-224 in NSCLC cells co-culture with CAFs for 24 hours were detected by RT-PCR. (D) The proliferation of NSCLC cells co-culture with CAFs for 24 hours was determined by the CCK8 assay. (E) Colony formation assay was used to evaluate the cell colony of NSCLC cells co-culture with CAFs for 24 hours. (F) Transwell assay was used to test the invasive ability of NSCLC cells co-culture with CAFs for 24 hours. (G) EMT markers including E-cadherin, Vimentin, and N-cadherin of NSCLC cells co-culture with CAFs for 24 hours were determined by western blot. (H) Tube formation assay was conducted to monitor the tube formation ability of HUVECs co-culture with CAFs for 24 hours. *, ** and *** represents P<0.05, P<0.01 and P<0.001, respectively. N=3.