Study on the Inhibitory Effect of Small Interfering RNA on Yes-Associated Protein Expression in Human Hepatoma Cell Lines
Suqin Zhang
Abstract
Suqin Zhang
Abstract
Objective:To investigate the inhibitory effect of small interfering RNA(siRNA)on yes-associated protein(YAP)expression and adriamycin(ADM)chemosensitivity of hepatoma cell lines.Methods:The siRNA sequences were designed based on YAP gene and were transfected into four different kinds of hepatoma cells(PLC/PRF/5,Huh-7,MHCC97 H,MHCC97L)through lipofectamineTM2000.The inhibition result of YAP expression in hepatoma cells was measured by Western blotting and fluorescence microscope.The viability rate of hepatoma cells was measured by methyl thiazolyl tetrazolium(MTT)colorimetry after treating with different concentrations of ADM.Results:The siRNA expression vector to aim directly at YAP gene was constructed successfully.Western blotting revealed that transfection with siRNA could inhibit the expression of YAP protein in hepatoma cell lines.The expression of YAP protein in hepatoma cell lines was specially and effectually inhibited by RNA interference(RNAi).MTT colorimetry detection result revealed that the viability rate of hepatoma cell lines significantly decreased after transfection with YAP siRNA.Conclusions:Inhibiting the expression of YAP by siRNA can enhance the sensitivity of hepatoma cell lines to ADM.
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Objective:To investigate the inhibitory effect of small interfering RNA(siRNA)on yes-associated protein(YAP)expression and adriamycin(ADM)chemosensitivity of hepatoma cell lines.Methods:The siRNA sequences were designed based on YAP gene and were transfected into four different kinds of hepatoma cells(PLC/PRF/5,Huh-7,MHCC97 H,MHCC97L)through lipofectamineTM2000.The inhibition result of YAP expression in hepatoma cells was measured by Western blotting and fluorescence microscope.The viability rate of hepatoma cells was measured by methyl thiazolyl tetrazolium(MTT)colorimetry after treating with different concentrations of ADM.Results:The siRNA expression vector to aim directly at YAP gene was constructed successfully.Western blotting revealed that transfection with siRNA could inhibit the expression of YAP protein in hepatoma cell lines.The expression of YAP protein in hepatoma cell lines was specially and effectually inhibited by RNA interference(RNAi).MTT colorimetry detection result revealed that the viability rate of hepatoma cell lines significantly decreased after transfection with YAP siRNA.Conclusions:Inhibiting the expression of YAP by siRNA can enhance the sensitivity of hepatoma cell lines to ADM.
Key concepts: Transfection, Small interfering RNA, RNA interference, Molecular biology, Blot, Viability assay, Cell culture, MTT assay