Knock-down of amphiregulin inhibits cellular invasion in inflammatory breast cancer.
Knock-down of amphiregulin inhibits cellular invasion in inflammatory breast cancer.
复制标题
DOI：
10.1002/jcp.22620
复制
发表时间：
2011-10
影响因子：
5.6
通讯作者：
Ethier, Stephen P.
中科院分区：
文献类型：
--
作者：
Baillo, Andrea;Giroux, Craig;Ethier, Stephen P.
我们之前已经表明，SUM-149人乳腺癌细胞需要AREG/EGFR自分泌环进行细胞增殖。我们还证明了AREG可以增加EGFR的稳定性并促进EGFR定位于质膜。在本研究中，我们通过慢病毒感染AREG shRNA成功地敲低了SUM-149细胞中的AREG表达。在AREG表达不存在的情况下，SUM-149细胞生长减慢，但未完全抑制。此外，用AREG shRNA构建体感染的细胞通过蛋白质印迹显示EGFR蛋白表达增加。免疫荧光和共聚焦显微镜显示，AREG敲低后，EGFR继续定位于细胞表面。软琼脂试验表明，AREG敲低细胞保留锚定非依赖性生长能力。另外，乳腺球形成测定和Adefluor染色分析显示AREG表达的敲低不影响干细胞表型的表达。然而，在AREG敲低后，SUM-149细胞表现出其侵入基质胶基质的能力显著降低。与该观察结果一致，微阵列分析比较了用非沉默载体感染的细胞与AREG敲低细胞，鉴定了与侵袭性表型相关的基因，如RHOB和DKK 1，以及与细胞运动相关的网络，如整合素连接的激酶信号传导和粘着斑激酶信号传导。还发现AREG调节这些细胞中的WNT和Notch信号传导。因此，AREG在调节侵袭性表型中起作用，并且我们提出这种调节可能是通过当AREG激活质膜定位的EGFR时发生的改变的信号传导。
We have previously shown that SUM-149 human breast cancer cells require an AREG/EGFR autocrine loop for cell proliferation. We also demonstrated that AREG can increase EGFR stability and promote EGFR localization to the plasma membrane. In the present studies we successfully knocked-down AREG expression in SUM-149 cells by lenti-viral infection of AREG shRNA. In the absence of AREG expression, SUM-149 cell growth was slowed, but not completely inhibited. Furthermore, cells infected with AREG shRNA constructs showed an increase in EGFR protein expression by western blot. Immunofluorescence and confocal microscopy showed that following AREG knock-down, EGFR continued to localize to the cell surface. Soft agar assays demonstrated that AREG knock-down cells retain anchorage-independent growth capacity. Additionally mammosphere forming assays and Adefluor staining analysis showed that knock-down of AREG expression did not affect the expression of stem cell phenotypes. However, following AREG knock-down, SUM-149 cells demonstrated a dramatic decrease in their ability to invade a Matrigel matrix. Consistent with this observation, microarray analysis comparing cells infected with a non-silencing vector to the AREG knock-down cells, identified genes associated with the invasive phenotype such as RHOB and DKK1, and networks associated with cell motility such as integrin-linked kinase signaling, and focal adhesion kinase signaling. AREG was also found to modulate WNT and Notch signaling in these cells. Thus, AREG functions in regulating the invasive phenotype, and we propose that this regulation may be through altered signaling that occurs when AREG activates plasma membrane localized EGFR.
登录
查看更多内容
影响因子：
11.2
作者：
Charafe-Jauffret E;Ginestier C;Iovino F;Wicinski J;Cervera N;Finetti P;Hur MH;Diebel ME;Monville F;Dutcher J;Brown M;Viens P;Xerri L;Bertucci F;Stassi G;Dontu G;Birnbaum D;Wicha MS
通讯作者：
Wicha MS
影响因子：
8.4
作者：
Price, JT;Wilson, HM;Haites, NE
通讯作者：
Haites, NE
影响因子：
11.4
作者：
Gschwind, A;Hart, S;Ullrich, A
通讯作者：
Ullrich, A
影响因子：
11.5
作者：
Mazieres, J;Antonia, T;Favre, G
通讯作者：
Favre, G
影响因子：
6.4
作者：
OZAWA, S;UEDA, M;SHIMIZU, N
通讯作者：
SHIMIZU, N