shRNA干扰沉默RNPC1a基因对SUM1315细胞增殖、迁移及侵袭的影响
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国家自然科学基金资助(81272916)


Effect of shRNA interference targeting RNPC1a gene on proliferation,migration and invasion of cell line SUM1315
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    摘要:

    目的:研究RNPC1a基因对SUM1315细胞增殖-迁移及侵袭的影响。方法:应用脂质体技术将RNPC1a 短发夹RNA(shRNA)转染至SUM1315细胞,通过嘌呤霉素筛选出稳定细胞株,采用实时荧光定量PCR和Western blot方法检测其干涉效率,并用噻唑蓝(MTT)比色法检测细胞的增殖情况,划痕实验及Transwell细胞侵袭实验检测其迁移和侵袭能力。结果:经qRT-PCR和Western blot方法证实稳定转染RNPC1a shRNA的SUM1315细胞中RNPC1a的mRNA和蛋白水平较对照组均明显下降(P < 0.05),同时RNPC1a干扰后细胞的增殖-迁移及侵袭能力均减弱(P < 0.05)。结论:RNPC1a shRNA能有效地降低SUM1315细胞中RNPC1a基因的表达,RNPC1a基因沉默可以抑制SUM1315细胞的增殖-迁移及侵袭。

    Abstract:

    Objective:To investigate inhibitory effect of RNPC1a short hairpin RNA (shRNA) on the expression of RNPC1a in cell line SUM1315 and determine the effect of RNPC1a down-expression on the proliferation,migration and invasion of SUM1315 cells. Methods:The RNPC1a shRNA was transfected into SUM1315 cells with lipofectine. Puromycin was used for selecting stable cell line. Real-time PCR and Western blot were performed to analyze the mRNA and protein expression of RNPC1a in SUM1315 cells. Cell proliferation,migration and invasion were assessed by methyl thiazol tetrazolium(MTT),wound-healing experiment and Matrigel invasion assay. Results:The mRNA and protein expressions of RNPC1a gene in the shRNA-RNPC1a-SUM1315 group were significantly lower than those in the control group,confirmed by RT-PCR and Western blot,respectively(P < 0.05). The proliferation of SUM1315 cells was markedly inhibited by RNPC1a shRNA with the inhibition rate at 26.5% (P < 0.05),while scratch repair time extended. Meantime transwell cell invasion assay showed that the invasive ability of SUM1315 cell was also inhibited in the shRNA-RNPC1a-SUM1315 group(23 ± 7),compared with the SUM1315 group(213 ± 12)(P < 0.01). Conclusion:Down-regulation of RNPC1a by RNPC1a shRNA can inhibit the proliferation,migration and invasion of SUM1315 cells.

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梁秀清,潘 红,周文斌,薛金秋,成 琳,王 莹,丁 强. shRNA干扰沉默RNPC1a基因对SUM1315细胞增殖、迁移及侵袭的影响[J].南京医科大学学报(自然科学版),2013,(7):861-866

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  • 收稿日期:2013-01-06
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  • 在线发布日期: 2013-07-09
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