Objective:To develop an engineered therapeutic tool based on miR-214,analyze the effects of regulating miR-214 on the biological behavior of cervical cancer cells,and explore its underlying mechanisms.Methods:Two type...Objective:To develop an engineered therapeutic tool based on miR-214,analyze the effects of regulating miR-214 on the biological behavior of cervical cancer cells,and explore its underlying mechanisms.Methods:Two types of miR-214 nano-paper carriers were self-designed according to different ratios.The stability,cellular uptake rate,and targeting ability of different carriers were tested.Human cervical epithelial cells and C6330 He La human cervical cancer cells were cultured.The human cervical epithelial cells in the logarithmic growth phase were used as the blank group.The C6330 He La human cervical cancer cells in the logarithmic growth phase were randomly divided into five groups.Cells from each group were seeded in cell culture plates.The blank group and the model group received no treatment.The miR-214 inhibition group was transfected with miR-214 inhibitor.The miR-214 activation group was delivered miR-214 by high-performance nano-lipid carriers.The MEK3 knockout group was transfected with miR-214 mimic after MEK3 gene knockout.The relative expression levels of miR-214 in each group of cells at 24,48,and 72 h after transfection were detected by real-time fluorescence quantitative PCR.The cell proliferation ability was evaluated by measuring the absorbance(OD)value using the MTT colorimetric method.The cell migration ability was evaluated by the cell scratch assay,and the cell apoptosis rate was evaluated by cell apoptosis detection.Results:The particle size of the 4∶5∶1 ratio carrier was significantly smaller than that of the 7∶2∶1 ratio carrier,while the Zeta potential,encapsulation efficiency,fluorescence intensity,and relative expression level of miR-214 were significantly higher than those of the 7∶2∶1 ratio carrier(P<0.05).After miR-214 inhibition and activation treatment in cervical cancer cells,the relative expression levels of miR-214 in cervical cancer cells decreased and increased respectively,and MEK3 knockout did not affect the change in the relative expression level of miR-214.The OD value of the miR-214 inhibition group was significantly higher than that of the model group,and the OD values of the miR-214 activation group and the MEK3 knockout group were significantly lower than that of the model group(P<0.05).The OD value of the miR-214 activation group was significantly lower than that of the MEK3 knockout group(P<0.05).After miR-214 inhibition treatment in cervical cancer cells,the cell migration ability was significantly enhanced,and the cell apoptosis rate was significantly reduced.After miR-214 activation treatment,the cell migration ability was significantly reduced,and the cell apoptosis rate was significantly increased.However,after knocking out the MEK3 gene,the cell proliferation,migration,and apoptosis abilities all decreased.Conclusion:Activating miR-214 can positively affect the biological behavior of cervical cancer cells.In this study,a nano-paper carrier was self-designed through engineering means,which can achieve efficient delivery of miR-214 and provide a new technological platform for the treatment of cervical cancer.展开更多
文摘Objective:To develop an engineered therapeutic tool based on miR-214,analyze the effects of regulating miR-214 on the biological behavior of cervical cancer cells,and explore its underlying mechanisms.Methods:Two types of miR-214 nano-paper carriers were self-designed according to different ratios.The stability,cellular uptake rate,and targeting ability of different carriers were tested.Human cervical epithelial cells and C6330 He La human cervical cancer cells were cultured.The human cervical epithelial cells in the logarithmic growth phase were used as the blank group.The C6330 He La human cervical cancer cells in the logarithmic growth phase were randomly divided into five groups.Cells from each group were seeded in cell culture plates.The blank group and the model group received no treatment.The miR-214 inhibition group was transfected with miR-214 inhibitor.The miR-214 activation group was delivered miR-214 by high-performance nano-lipid carriers.The MEK3 knockout group was transfected with miR-214 mimic after MEK3 gene knockout.The relative expression levels of miR-214 in each group of cells at 24,48,and 72 h after transfection were detected by real-time fluorescence quantitative PCR.The cell proliferation ability was evaluated by measuring the absorbance(OD)value using the MTT colorimetric method.The cell migration ability was evaluated by the cell scratch assay,and the cell apoptosis rate was evaluated by cell apoptosis detection.Results:The particle size of the 4∶5∶1 ratio carrier was significantly smaller than that of the 7∶2∶1 ratio carrier,while the Zeta potential,encapsulation efficiency,fluorescence intensity,and relative expression level of miR-214 were significantly higher than those of the 7∶2∶1 ratio carrier(P<0.05).After miR-214 inhibition and activation treatment in cervical cancer cells,the relative expression levels of miR-214 in cervical cancer cells decreased and increased respectively,and MEK3 knockout did not affect the change in the relative expression level of miR-214.The OD value of the miR-214 inhibition group was significantly higher than that of the model group,and the OD values of the miR-214 activation group and the MEK3 knockout group were significantly lower than that of the model group(P<0.05).The OD value of the miR-214 activation group was significantly lower than that of the MEK3 knockout group(P<0.05).After miR-214 inhibition treatment in cervical cancer cells,the cell migration ability was significantly enhanced,and the cell apoptosis rate was significantly reduced.After miR-214 activation treatment,the cell migration ability was significantly reduced,and the cell apoptosis rate was significantly increased.However,after knocking out the MEK3 gene,the cell proliferation,migration,and apoptosis abilities all decreased.Conclusion:Activating miR-214 can positively affect the biological behavior of cervical cancer cells.In this study,a nano-paper carrier was self-designed through engineering means,which can achieve efficient delivery of miR-214 and provide a new technological platform for the treatment of cervical cancer.
