Biotech Research and Innovation Center (BRIC), University of Copenhagen, Denmark.
Weizmann Institute of Science, Rehovot, Israel.
FEBS Open Bio. 2021 Jun;11(6):1579-1592. doi: 10.1002/2211-5463.13149. Epub 2021 May 2.
Activated Cdc42-associated kinase 1 (ACK1), a widely expressed nonreceptor tyrosine kinase, is often amplified in cancer and has been shown to interact with Cell division cycle 42 (Cdc42), Epidermal growth factor receptor (EGFR), and several other cancer-relevant molecules, suggesting a possible role for ACK1 in development and tumor formation. To directly address this scenario, we generated mice lacking a functional ACK1 gene (ACK1 ko) using CRISPR genome editing. ACK1 ko mice developed normally, displayed no obvious defect in tissue maintenance, and were fertile. Primary ACK1-null keratinocytes showed normal phosphorylation of EGFR, but a tendency toward reduced activation of AKT serine/threonine kinase 1 (Akt) and Mitogen-activated protein kinase 1 (Erk). DMBA/TPA-induced skin tumor formation did not reveal significant differences between ACK1 ko and control mice. Deletion of the ACK1 gene in the breast cancer cell lines MDA-MB-231, 67NR, MCF7, 4T1, and T47D caused no differences in growth. Furthermore, EGF-induced phosphorylation kinetics of Erk, Akt, and p130Cas were not detectably altered in T47D cells by the loss of ACK1. Finally, loss of ACK1 in MDA-MB-231 and T47D breast cancer cells had a very limited or no effect on directed cell migration. These data do not support a major role for ACK1 in Cdc42 and EGFR signaling, development, or tumor formation.
激活的 Cdc42 相关激酶 1(ACK1)是一种广泛表达的非受体酪氨酸激酶,通常在癌症中扩增,并已被证明与细胞分裂周期 42(Cdc42)、表皮生长因子受体(EGFR)和其他几种与癌症相关的分子相互作用,这表明 ACK1 在发育和肿瘤形成中可能发挥作用。为了直接解决这一情况,我们使用 CRISPR 基因组编辑生成了缺乏功能性 ACK1 基因(ACK1 ko)的小鼠。ACK1 ko 小鼠正常发育,组织维持无明显缺陷,且具有生育能力。原代 ACK1 缺失的角质形成细胞显示 EGFR 的磷酸化正常,但 AKT 丝氨酸/苏氨酸激酶 1(Akt)和丝裂原活化蛋白激酶 1(Erk)的激活趋势降低。DMBA/TPA 诱导的皮肤肿瘤形成在 ACK1 ko 和对照小鼠之间没有明显差异。乳腺癌细胞系 MDA-MB-231、67NR、MCF7、4T1 和 T47D 中 ACK1 基因的缺失对生长没有差异。此外,ACK1 缺失不会改变 T47D 细胞中 EGF 诱导的 Erk、Akt 和 p130Cas 的磷酸化动力学。最后,在 MDA-MB-231 和 T47D 乳腺癌细胞中缺失 ACK1 对定向细胞迁移的影响非常有限或没有。这些数据不支持 ACK1 在 Cdc42 和 EGFR 信号转导、发育或肿瘤形成中发挥主要作用。