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Knockdown of p57 gene inhibits breast cancer cell proliferation

机译:P57基因的敲低抑制乳腺癌细胞增殖

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摘要

The aim of the study was to investigate possible effects of p57 on the growth of the human MCF-7 and rat SHZ-88 breast cancer cell lines. Specific oligonucleotide sequences containing small hairpin structure were inserted into a small interfering RNA (siRNA) expression vector. The human MCF-7 and rat SHZ-88 breast cancer cell lines were transfected with recombinant plasmids. The p57 gene expression was blocked in the human MCF-7 breast and rat SHZ-88 breast cancer cells, using chemically modified siRNA. The p57 expression level was evaluated using quantitative polymerase chain reaction (qPCR) and western blot analysis. Immunofluorescence was conducted to detect p57 expression in the breast cancer cells. Tetrazolium blue (MTT) method was employed to detect the effect of p57 inhibition on the proliferation of the MCF-7 and SHZ-88 cell lines. Cell proliferation in the experimental group was significantly reduced. Immunofluorescence assay results showed p57 siRNA effectively inhibited the p57 level in the MCF-7 and SHZ-88 cells. RT-PCR results showed that 48 h after transfection, the p57 mRNA level in the transfected group was significantly lower compared with the control group. In conclusion, p57 effectively inhibited the proliferation of breast cancer after stable interference.
机译:该研究的目的是研究P57对人MCF-7和大鼠SHZ-88乳腺癌细胞系生长的可能影响。将含有小发夹结构的特异性寡核苷酸序列插入小干扰RNA(siRNA)表达载体中。用重组质粒转染人MCF-7和大鼠SHZ-88乳腺癌细胞系。使用化学改性的siRNA,在人MCF-7乳腺癌和大鼠SHZ-88乳腺癌细胞中封闭P57基因表达。使用定量聚合酶链反应(QPCR)和Western印迹分析评估P57表达水平。进行免疫荧光以检测乳腺癌细胞中的p57表达。使用四唑鎓(MTT)方法检测p57抑制对MCF-7和SHZ-88细胞系的增殖的影响。实验组细胞增殖显着降低。免疫荧光测定结果显示P57 siRNA在MCF-7和SHZ-88细胞中有效地抑制P57水平。 RT-PCR结果表明,转染后48小时,转染基团的P57 mRNA水平与对照组相比显着降低。总之,P57在稳定干扰后有效抑制乳腺癌的增殖。

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