RESUMO
AIM: To observe the effects of UO-126 on the expression of F-box and WD repeat domain-containing protein 7(FBW7)and on the proliferation of human cervical cancer cell lines (HeLa cells). METHODS: HeLa cells were treated with different concentrations of UO-126, MTT assay was used to observe the proliferation of HeLa cells. Immunofluorescence showed the location and expression of FBW7 in HeLa cells. The mRNA and protein expression of FBW7 were detected by RT-PCR and Western blot before and after mitogen-activated protein kinases (MAPK)signal was blocked by UO-126 a MAPK inhibitor. RESULTS: MTT results showed that the concentration range of MAPK signaling pathway inhibitor UO-126 inhibited the proliferation of HeLa cells in a concentration-and time-dependent manner(P<0.05). Immunofluorescence showed that the expression of positive FBW7 had increased after HeLa cells were treated with UO-126. RT-PCR and Western blot exhibited that the FBW7 mRNA and protein expression had significantly increased before and after HeLa cells were treated with UO-126(P<0.05). CONCLUSION: UO-126 could inhibit HeLa cells proliferation, FBW7 lied downstream of MAPK signaling pathway.
Assuntos
Butadienos/farmacologia , Proteínas de Ciclo Celular/genética , Proteínas F-Box/genética , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Nitrilas/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Ubiquitina-Proteína Ligases/genética , Western Blotting , Proteínas de Ciclo Celular/análise , Proliferação de Células/efeitos dos fármacos , Proteínas F-Box/análise , Proteína 7 com Repetições F-Box-WD , Imunofluorescência , Células HeLa , Humanos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ubiquitina-Proteína Ligases/análiseRESUMO
AIM: To study the anti-tumor effects and mechanisms of cisplatin(DDP) combined with exosomes. METHODS: The cell-growth inhibition of DDP on H(22); cells was analyzed by MTT assay.The mice were immunized by H(22); cell-derived exosomes. The exosomes elicited CTL-specific cytotoxicity and DDP enhanced cytotoxicity in combination with CTL were detected by (3)H-TdR release assay. The effect of DDP on mRNA and protein levels of Fas in H(22); was analyzed using RT-PCR and Western blot.The expression of FasL on spleen lymphocyte was determined by RT-PCR. BALB/c mice inoculated with hepatoma carcinoma cell line H(22); were used as tumor models.The mice received exosomes solely or in combination with DDP.The survival of the mice was observed. RESULTS: DDP inhibited H(22); cell in a dose-dependant manner. The exosomes elicited CTL-specific cytotoxicity was enhanced by DDP (P<0.05). RT-PCR and Western blot showed Fas increased gradually after administering DDP on H(22); cells.RT-PCR also indicated the mRNA level of FasL on mice spleen lymphocyte increased after immunized by exosomes. Compared with other groups, the combination group(DDP plus exosomes)could statistically prolong the survival time (P<0.05). CONCLUSION: The therapy of DDP combined with exosomes had significant synergistic effect against tumor. The mechanism of synergistic effect includes enhancement of CTL activity.
