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机构地区:[1]武警辽宁总队医院泌尿外科,辽宁沈阳110034
出 处:《武警医学院学报》2011年第10期782-785,F0003,共5页Acta Academiae Medicinae CPAPF
摘 要:【目的】通过pSilencer4.1-cmv-TrkC siRNA干涉质粒转染下调人前列腺癌细胞系PC3中酪氨酸激酶受体C(tyrosinelcinase C,TrkC)的表达,观察TrkC在前列腺癌细胞中的作用。【方法】根据TrkC基因序列设计并合成siRNA,构建质粒。用脂质体将pSilencer/TrkC和对照空载体pSilencer转染人前列腺癌细胞系PC3,通过Western blotting检测转染细胞的TrkC表达情况,绘制细胞生长曲线,流式细胞仪检测TrkC对细胞周期及凋亡的影响。【结果】成功利用脂质体转染PC3细胞后,Western blotting鉴定结果显示:TrkC的表达水平显著降低,细胞生长曲线及流式细胞仪检测结果显示pSilencer4.1-cmv-TrkC脂质体转染的PC3细胞出现生长抑制及凋亡增加。【结论】下调TrkC表达可以抑制前列腺癌细胞PC3增殖,TrkC可能参与了前列腺癌的发生发展过程。【Objective】 To observe the effect of tyrosine kinase C(TrkC) in prostate cancer cell line(PC3) by siRNA interference technology.【Methods】Using LipofectAmineTM2000 reagent,the human prostate cancer cell line PC3 was transfectted with siRNA eukaryotic expression vectors pSilencer/TrkC and contro1 plasmid pSilencer.Western blotting was used to testify the protein level in the transfected and parental PC3 cells.Cell growth curves and flow cytometry(FCM) were used to detected the function of TrkC mediated by pSilencer4.1-cmv in PC3.【Results】Psilencer/TrkC or pSilencer was stably transfectted into prostate cancer PC3 cells using LipofectAmineTM 2000 reagent.Western blotting showed that expression of protein significantly decreased in the pSilencer/TrkCs transfectted PC3 cells.While in control plasmid pSilencer transfectted PC3 cells,TrkC expression remained in the same level as the parental cells.Cell growth curves and FCM showed that the growth of PC3 cells was inhibited in the experimental groups as compared with the control group.【Conclusion】Down-regulation of TrkC could inhibit the proliferation of PC3 cells,and TrkC may play an important role in the tumorigenesis and development of PCA.
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