【摘 要】
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To investigate the effect of sequence-specific small interfering RNA (siRNA)on suppressing cyclinD 1 expression and proliferation and cell cycle and expression of related signal transduction pathway o
【机 构】
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the First Clinical College of Harbin Medical University, Harbin,Heilongjiang150001,China
【出 处】
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第四届中国临床微生物学大会暨微生物学与免疫学论坛
论文部分内容阅读
To investigate the effect of sequence-specific small interfering RNA (siRNA)on suppressing cyclinD 1 expression and proliferation and cell cycle and expression of related signal transduction pathway of fibroblasts derived from keloid(KFB).The plasmid expression vector of siRNA targeted against cyclinD1 was constructed and transfected into KFB with LipofcctamincTM2000.The effect of sequence specific siRNA in suppressing the proliferation of KFB were detected by MTT assay.Flow cytometry(FCM) were used for evaluation of cell cycle.The changes of cyclinD 1 expression were detected by fluorescent quantitative PCR (FQ-PCR) methods.The expression of cyliinDI, cyclinE, CDK2, CDK4, P27 and pRb were detected by and Western blot and immunohistochemical method.The sequence-specific siRNA effectively suppressed cyclinD1 expression at both mRNA and protein levels with inhibition rate of 92.83% and 72.53% (P<0.01).Significantly inhibited the proliferation of KFB, and changed cell cycle in percentage of G1 phase cells was increased and G2/M phase decreased compared with the controls groups KFB(P<0.05).The after transfection, decreased the expression ofCDK2, CDK4 and pRb, enhance the expression of P16 and P27 (P<0.05).The plasmid expression vector for the RNA i against cyclinD1 constructed in the study can effectively and specifically suppress cyclinD1 expression, and progression of G I/S is effect by G1 phase related regulatory protein ,and suppresses proliferation of KFB.
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