RNAi-mediated knockdown of FANCF suppresses cell proliferation, migration, invasion, and drug resistance potential of breast cancer cells.
RNAi-mediated knockdown of FANCF suppresses cell proliferation, migration, invasion, and drug resistance potential of breast cancer cells.
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DOI：
10.1590/1414-431x20132938
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发表时间：
2014-01
期刊：
影响因子：
--
通讯作者：
Wei MJ
中科院分区：
文献类型：
--
作者：
Zhao L;Li N;Yu JK;Tang HT;Li YL;He M;Yu ZJ;Bai XF;Zheng ZH;Wang EH;Wei MJ
Fanconi贫血互补组F蛋白（FANCF）是维持FA/BRCA这一DNA损伤反应通路功能的关键因子。然而，FANCF在乳腺癌中的功能作用尚未阐明。我们在体外进行了内源性FANCF的特异性FANCF-shRNA敲低。用CCK-8测定法测量细胞活力。用碱性彗星试验评估DNA损伤。流式细胞仪检测细胞凋亡、细胞周期和药物蓄积。蛋白质表达水平通过使用特异性抗体的Western印迹测定。基于这些结果，我们使用细胞迁移和侵袭试验来证明FANCF在这些过程中的关键作用。FANCF shRNA能有效抑制FANCF的表达。我们发现FANCF基因敲减的乳腺癌细胞（MCF-7和MDA-MB-435S）的增殖被显著抑制，细胞周期停滞在S期，诱导凋亡和DNA片段化。FANCF的抑制也导致细胞迁移和侵袭减少。此外，FANCF敲低增强了乳腺癌细胞对阿霉素的敏感性。这些结果表明，FANCF可能是乳腺癌分子治疗干预的潜在靶点。
Fanconi anemia complementation group F protein (FANCF) is a key factor, which maintains the function of FA/BRCA, a DNA damage response pathway. However, the functional role of FANCF in breast cancer has not been elucidated. We performed a specific FANCF-shRNA knockdown of endogenous FANCF in vitro. Cell viability was measured with a CCK-8 assay. DNA damage was assessed with an alkaline comet assay. Apoptosis, cell cycle, and drug accumulation were measured by flow cytometry. The expression levels of protein were determined by Western blot using specific antibodies. Based on these results, we used cell migration and invasion assays to demonstrate a crucial role for FANCF in those processes. FANCF shRNA effectively inhibited expression of FANCF. We found that proliferation of FANCF knockdown breast cancer cells (MCF-7 and MDA-MB-435S) was significantly inhibited, with cell cycle arrest in the S phase, induction of apoptosis, and DNA fragmentation. Inhibition of FANCF also resulted in decreased cell migration and invasion. In addition, FANCF knockdown enhanced sensitivity to doxorubicin in breast cancer cells. These results suggest that FANCF may be a potential target for molecular, therapeutic intervention in breast cancer.