ABSTRACT
Objective: To use autophagy inhibitors combined with radiation to treat the oral squamous cell carcinoma CAL-27 and KB cells,and to explore the influence of autophagy in the oral cancer radiation sensitivity and its mechanisms. Methods:The human oral squamous cell carcinoma CAL-27 and KB cells were divided into control group,CQ group,3-MA group,IR group,CQ+IR group,and 3-MA+IR group. The survival rate was detected by MTT method and the autophagy of CAL-27 cells was observed by immunofluorescence method and laser scanning confocal microscope.The expression levels of LC3 and beclin-1 were detected by Western blotting method. The apoptotic rate was determined by flow cytometry with Annexin Ⅴ/PI doulde staining. Results:Compared with IR group,the survival rates in 3-MA + IR and CQ+ IR groups were signifcantly decreased (P < 0.05 ).The autophagy levels of cells in IR group were significantly higher than those in control group, CQ group, 3-MA group,CQ+IR group,and 3-MA+IR group (P <0.05).The expression levels of LC3 and beclin-1 proteins in IR group were significantly higher than those in control group,CQ+ IR group,and 3-MA+ IR group (P < 0.05). The apoptotic rates in IR,3-MA+ IR,and CQ+ IR groups were markedly higher than those in control group. Compared with IR group,the apoptotic rates in CQ+IR and 3-MA+IR groups were significantly increased (P <0.05).Conclusion:Radiotherapy can induce the increase of autophagy level of oral squamous cell carcinoma cells. Inhibiors of autophagy can increase the radio-sensitivity of oral squamous cell carcinoma cells by inhibiting the proliferation and inducing the apoptosis.
ABSTRACT
Objective:To investigate the effect of poly (lactic acid)(PLA)electrospun membranes loaded with cisplatin and chloroquine on the oral squamous cell carcinoma CAL-27 cells,and to explore the method to prevent the recurrence of oral cancer.Methods: The DDP/PLA membranes, CQ/DDP/CQ/PLA membranes and CQ/DDP/PLA membranes were prepared by electrospinning.Then the micro morphology of three kinds of membranes were observed by scanning electron microscope (SEM);the degradation rate of PLA membrane was measuredby UV spectrophotometric.The LC3-Ⅱ expression level in CAL-27 cells was detected by laser scanning confocal microscope.The survival rate of CAL-27 cells was detected by MTT method.Results:The SEM results showed that the nanofibers of DDP/PLA,CQ/DDP/PLA and CQ/DDP/CQ/PLA membranes were continuous and smooth with uniform diameters.The degrated time of membranes was about 21 d.The MTT result showed that compared with control group,at first,the effects of cell killing of DDP/PLA membranes,CQ/DDP/CQ/PLA membranes and CQ/DDP/PLA membranes were not obvious;as the extension of time,the survival rates of CAL-27 cells in DDP/PLA membranes group,CQ/DDP/CQ/PLA membranes group and CQ/DDP/PLA membranes group were decreased (P <0.05).The immunofluorescence results showed that the fluorescence intensity of LC3-Ⅱ in CQ/DDP/CQ/PLA membranes group and CQ/DDP/PLA membranes group were lower than that in DDP/PLA membranes group.Conclusion:CQ/DDP/PLA membranes with sustained-release effect can increase the sensitivity of CAL-27 cells to DDP and enchance the killer effect of DDP on the CAL-27 cells.