反义EGFR寡核苷酸对AngⅡ促血管平滑肌细胞增殖效应的影响

Effect of antisense senquences of epidermal growth factor receptor on proliferation of vascular smooth muscle cells activated by AngⅡ

目的探讨脂质体介导的反义表皮生长因子受体(EGFR)寡核苷酸基因转染对血管紧张素Ⅱ(AngⅡ)促大鼠血管平滑肌细胞(VSMC)增殖的影响。方法用EGFR寡核苷酸脂质体复合物转染Sprague-Dawley大鼠VSMC,通过RT-PCR、Western-Bloting分别检测转染后EGFR mRNA及蛋白的表达情况,用3H-Tdr掺入法检测经EGFR寡核苷酸转染再用AngⅡ刺激VSMC的增殖情况。结果反义EGFR寡核苷酸转染大鼠VSMC后,EGFR mRNA及蛋白的表达较正义组及对照组显著减少(P<0.01);用AngⅡ刺激后,反义组细胞的3H-Tdr掺入较正义组及对照组显著降低(P<0.01)。结论脂质体介导的反义EGFR寡核苷酸转染可以减弱AngⅡ的促VSMC增殖效应。

AIM To investigate the effect of antisense sequences of epidermal growth factor receptor （EGFR） on the proliferation of vascular smooth muscle cells （VSMCs） activated by angiotensin Ⅱ （ Ang Ⅱ ）. METHODS Sprague-Dawley rat VSMCs were transfected with antisense sequences of EGFR and stimulated by Ang Ⅱ. EGFR mRNA and protein in VSMCs were detected by reverse transcription polymerase chain reaction （RT-PCR） and Western-Bloting. Proliferation of VSMCs was detected by ^3H-Tdr incorporation. RESULTS EGFR mRNA and protein in VSMCs transfected with antisense sequences of EGFR were significantly lower than those transfected with sense sequences of EGFR and control （P 〈 0.01 ） . The ^3H-Tdr incorporation of VSMCs transfected with antisense senquences of EGFR was markedly lower than that transfected with sense sequences of EGFR and control（ P 〈 0.01 ）. CONCLUSION Antisense sequences of EGFR attenuate the proliferation of VSMCs activated by Ang Ⅱ , which indicates that EGFR plays an important role in the proliferation of VSMCs activated by Ang Ⅱ.