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The effect of small interfering RNA (siRNA) against the Bcl-2 gene on apoptosis and chemosensitivity in a canine mammary gland tumor cell line

机译:针对Bcl-2基因的小干扰RNA(siRNA)对犬乳腺肿瘤细胞株凋亡和化学敏感性的影响

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摘要

The aim of this study was to investigate whether downregulation of Bcl-2 expression by small interfering RNA (siRNA) against the canine Bcl-2 gene would enhance the apoptosis and sensitivity of a canine mammary gland tumor cell line (CF33) to doxorubicin. Transfections of CF33 with siRNA were performed using cationic liposomes. Sequence-specific downregulation of Bcl-2 expression was measured by semiquantitative RT-PCR and Western blot analysis. Total viable cells were determined by MTS assay and apoptotic cell rates were determined by the immunohistochemical analysis on ssDNA. Our data showed the siRNA downregulated Bcl-2 expression which increased apotosis and also increased the sensitivity of CF33 to doxorubicin. This study indicated that downregulation of Bcl-2 expression by siRNA would be useful as a new protocol to increase the effect of doxorubicin on treatment of canine mammary gland tumors, requiring a detailed evaluation of siRNA in vivo.
机译:这项研究的目的是调查小干扰RNA(siRNA)对犬Bcl-2基因的Bcl-2表达下调是否会增强犬乳腺肿瘤细胞系(CF33)对阿霉素的凋亡和敏感性。使用阳离子脂质体进行CF33与siRNA的转染。 Bcl-2表达的序列特异性下调是通过半定量RT-PCR和Western blot分析来测量的。通过MTS测定确定总活细胞,并通过对ssDNA的免疫组织化学分析确定凋亡细胞率。我们的数据显示,siRNA下调了Bcl-2表达,这增加了细胞凋亡,也增加了CF33对阿霉素的敏感性。这项研究表明,siRNA对Bcl-2表达的下调将作为增加阿霉素对犬乳腺肿瘤治疗作用的新方案是有用的,需要在体内对siRNA进行详细评估。

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