Lentiviral CRISPR/Cas9 nickase vector mediated BIRC5 editing inhibits epithelial to mesenchymal transition in ovarian cancer cells.
Lentiviral CRISPR/Cas9 nickase vector mediated BIRC5 editing inhibits epithelial to mesenchymal transition in ovarian cancer cells.
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DOI：
10.18632/oncotarget.21863
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发表时间：
2017-11-07
期刊：
影响因子：
--
通讯作者：
Yue J
中科院分区：
文献类型：
--
作者：
Zhao G;Wang Q;Gu Q;Qiang W;Wei JJ;Dong P;Watari H;Li W;Yue J
BIRC5编码survivin蛋白，是凋亡抑制家族的一员。Survivin在多种癌症中高表达，但在相应的正常组织中表达极低，其表达常与肿瘤转移和化疗耐药有关。我们报道survivin在卵巢癌中高度表达，并与患者总体生存率低密切相关。我们首次提供了survivin参与卵巢癌细胞上皮细胞向间充质细胞转化（EMT）的实验证据。在卵巢癌SKOV3和OVCAR3细胞中，慢病毒CRISPR/Cas9 nickase载体介导的BIRC5基因编辑通过上调上皮细胞标记物细胞角蛋白7和下调间充质标记物snail2、β-catenin和vimentin来抑制EMT。与这种分子方法一致，使用小分子survivin抑制剂YM155对卵巢癌细胞进行药理学治疗也抑制了这些卵巢癌细胞系的EMT。BIRC5的过表达促进了SKOV3细胞的EMT。通过分子或药理学方法，我们发现在这两种细胞系中，BIRC5破坏后细胞增殖、迁移和侵袭均受到显著抑制。抑制BIRC5表达也使细胞对紫杉醇治疗的反应增敏。此外，BIRC5表达的缺失减弱了SKOV3和OVCAR3细胞中的tgf - β信号传导。总的来说，我们的研究表明，破坏BIRC5表达通过减弱卵巢癌细胞中的tgf - β途径来抑制EMT。
BIRC5 encodes the protein survivin, a member of the inhibitor of apoptosis family. Survivin is highly expressed in a variety of cancers but has very low expression in the corresponding normal tissues, and its expression is often associated with tumor metastasis and chemoresistance. We report that survivin was highly expressed in ovarian cancer and strongly correlated with patient overall poor survival. For the first time, we provide experimental evidence that survivin is involved in epithelial to mesenchymal transition (EMT) in ovarian cancer cells. Lentiviral CRISPR/Cas9 nickase vector mediated BIRC5 gene editing led to the inhibition of EMT by upregulating epithelial cell marker, cytokeratin 7 and downregulating mesenchymal markers: snail2, β-catenin, and vimentin in both ovarian cancer SKOV3 and OVCAR3 cells. Consistent with this molecular approach, pharmacological treatment of ovarian cancer cells using a small molecule survivin inhibitor, YM155 also inhibited EMT in these ovarian cancer cell lines. Overexpression of BIRC5 promoted EMT in SKOV3 cells. Using molecular or pharmacological approaches, we found that cell proliferation, migration, and invasion were significantly inhibited following BIRC5 disruption in both cell lines. Inhibition of BIRC5 expression also sensitized cell responses to paclitaxel treatment. Moreover, loss of BIRC5 expression attenuated TGFβ signaling in both SKOV3 and OVCAR3 cells. Collectively, our studies demonstrated that disruption of BIRC5 expression inhibited EMT by attenuating the TGFβ pathway in ovarian cancer cells.
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