Research Paper Volume 12, Issue 8 pp 6756—6773

Exosomal miR-200c suppresses chemoresistance of docetaxel in tongue squamous cell carcinoma by suppressing TUBB3 and PPP2R1B

Figure 8. MiR-200c regulated docetaxel resistance in HSC-3 (HSC-3DR) cells via targeting TUBB3 and PPP2R1B. (A) Putative binding sites of miR-200c in 3’UTR of TUBB3 and PPP2R1B. (B) Relative luciferase activity was determined by luciferase reporter assays in HSC-3DR cells transfected with miR-200c mimics and luciferase vectors containing wild-type or mutant 3’UTR of TUBB3. (C) Relative luciferase activity was determined by luciferase reporter assays in HSC-3DR cells transfected with miR-200c mimics and luciferase vectors containing wild-type or mutant 3’UTR of PPP2R1B. (D) The mRNA expressions of TUBB3 and PPP2R1B were determined by qRT-PCR in HSC-3DR cells transfected with miR-200c-encoding lentiviral vectors (LV-200c). (E) The protein expressions of TUBB3 and PPP2R1B were determined by western blots in HSC-3DR cells transfected with miR-200c-encoding lentiviral vectors (LV-200c). (F) The protein expression of TUBB3 was determined by western blots in HSC-3DR cells transfected with TUBB3-carrying vectors with the wild-type or mutant miR-200c sequence. (G) The protein expression of PPP2R1B was determined by western blots in HSC-3DR cells transfected with PPP2R1B-carrying vectors with the wild-type or mutant miR-200c sequence. (H) Cell viability was determined by CCK8 assays in HSC-3DR cells transfected with TUBB3 or PPP2R1B-carrying vectors with the wild-type or mutant miR-200c sequence. (I) Apoptosis was determined by flow cytometry in HSC-3DR cells transfected with TUBB3 or PPP2R1B-carrying vectors with the wild-type or mutant miR-200c sequence. (J) Invasion ability was determined by Transwell assays in HSC-3DR cells transfected with TUBB3 or PPP2R1B-carrying vectors with the wild-type or mutant miR-200c sequence (scale bars = 50 μm). (K) Migration ability was determined by Transwell assays in HSC-3DR cells transfected with TUBB3 or PPP2R1B-carrying vectors with the wild-type or mutant miR-200c sequence (scale bars = 50 μm). Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.