Identification of WEE1 as a potential molecular target in cancer cells by RNAi screening of the human tyrosine kinome.
Identification of WEE1 as a potential molecular target in cancer cells by RNAi screening of the human tyrosine kinome.
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DOI：
10.1007/s10549-009-0571-2
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发表时间：
2010-07
影响因子：
3.8
通讯作者：
Lipkowitz, Stanley
中科院分区：
文献类型：
--
作者：
Murrow, Lyndsay M.;Garimella, Sireesha V.;Jones, Tamara L.;Caplen, Natasha J.;Lipkowitz, Stanley
乳腺癌可分为表达雌激素（ER）和孕激素（PR）受体的乳腺癌，具有ERBB 2（HER-2/Neu）扩增的乳腺癌，以及不表达ER、PR或ERBB 2扩增的乳腺癌（称为三阴性或基底样乳腺癌）。为了确定乳腺癌中潜在的分子靶点，我们对人酪氨酸激酶组进行了合成siRNA介导的RNAi筛选。在三阴性/基底样乳腺癌细胞系MDA-MB 231中进行的初步RNAi筛选，随后在该细胞系和另外两种三阴性/基底样乳腺癌细胞系BT 20和HCC 1937中进行的二次RNAi筛选和进一步研究，鉴定了G2/M检查点蛋白WEE 1作为潜在的治疗靶标。在所有乳腺癌亚型的细胞系中观察到对WEE 1抑制的相似敏感性。在乳腺癌细胞系中，RNAi介导的WEE 1沉默或小化合物抑制导致γ H2 AX水平增加，细胞周期S期停滞，细胞增殖显著降低。WEE 1抑制的细胞发生凋亡，如通过阳性膜联蛋白V染色、亚G1 DNA含量增加、凋亡形态、半胱天冬酶激活和泛半胱天冬酶抑制剂Z-VAD-FMK的拯救所证明的。相反，非转化的乳腺上皮细胞系MCF 10A在WEE 1沉默或抑制后没有表现出任何这些下游效应。这些结果确定WEE 1作为乳腺癌的潜在分子靶点。
Breast cancers can be classified into those that express the estrogen (ER) and progesterone (PR) receptors, those with ERBB2 (HER-2/Neu) amplification, and those without expression of ER, PR, or amplification of ERBB2 (referred to as triple-negative or basal-like breast cancer). In order to identify potential molecular targets in breast cancer, we performed a synthetic siRNA-mediated RNAi screen of the human tyrosine kinome. A primary RNAi screen conducted in the triple-negative/basal-like breast cancer cell line MDA-MB231 followed by secondary RNAi screens and further studies in this cell line and two additional triple-negative/basal-like breast cancer cell lines, BT20 and HCC1937, identified the G2/M checkpoint protein, WEE1, as a potential therapeutic target. Similar sensitivity to WEE1 inhibition was observed in cell lines from all subtypes of breast cancer. RNAi-mediated silencing or small compound inhibition of WEE1 in breast cancer cell lines resulted in an increase in γH2AX levels, arrest in the S-phase of the cell cycle, and a significant decrease in cell proliferation. WEE1-inhibited cells underwent apoptosis as demonstrated by positive Annexin V staining, increased sub-G1 DNA content, apoptotic morphology, caspase activation, and rescue by the pan-caspase inhibitor, Z-VAD-FMK. In contrast, the non-transformed mammary epithelial cell line, MCF10A, did not exhibit any of these downstream effects following WEE1 silencing or inhibition. These results identify WEE1 as a potential molecular target in breast cancer.
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影响因子：
3.8
作者：
Rahman, Monzur;Davis, Sean R.;Lipkowitz, Stanley
通讯作者：
Lipkowitz, Stanley
影响因子：
3.4
作者：
Johnston, CJ;Williams, JP;Finkelstein, JN
通讯作者：
Finkelstein, JN
影响因子：
50.3
作者：
Neve, Richard M.;Chin, Koei;Gray, Joe W.
通讯作者：
Gray, Joe W.
影响因子：
3.9
作者：
Petitjean, Audrey;Mathe, Ewy;Olivier, Magali
通讯作者：
Olivier, Magali
影响因子：
21.3
作者：
MacKeigan, JP;Murphy, LO;Blenis, J
通讯作者：
Blenis, J