Integrin β1 promotes gemcitabine resistance in pancreatic cancer through Cdc42 activation of PI3K p110β signaling
Integrin β1 promotes gemcitabine resistance in pancreatic cancer through Cdc42 activation of PI3K p110β signaling
复制标题
整合素 beta 1 通过 Cdc42 激活 PI3K p110 beta 信号传导促进胰腺癌吉西他滨耐药
DOI：
10.1016/j.bbrc.2018.09.061
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发表时间：
2018-10-20
影响因子：
3.1
通讯作者：
Cai, Qingping
中科院分区：
文献类型：
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作者：
Yang, Dejun;Tang, Yuan;Cai, Qingping
胰腺导管腺癌（PDAC）是最常见的恶性肿瘤之一，由于其对化疗的广泛耐药性，预后极差。我们以前的研究表明，整合素β 1表达上调PDAC和赋予吉西他滨耐药PDAC细胞通过信号通路，包括Cdc 42和AKT激活。但其确切的信号转导机制尚不清楚。在此，我们旨在阐明整合素01在PDAC中获得吉西他滨耐药的信号转导。从AsPC-1亲本细胞系（PCL）中选择耐药（DR）细胞。用蛋白质印迹法测定整合素β 1的表达。采用MTT法、切割型caspase-3免疫荧光法和Western blot法检测药物反应和敲低整合素β 1、Cdc 42或p110 β后磷脂酰肌醇3-激酶（PI 3 K）信号转导活性的变化。Western blot检测还检测了整合素01敲低后Cdc 42活性和p110 β表达的变化。用谷胱甘肽S-转移酶（GST）pull-down法检测Cdc 42与p110 β的相互作用。结果显示，整合素β 1在DR-AsPC-1细胞中表达上调，整合素β 1敲低显著降低整合素β 1的靶分子Cdc 42的活性和p110 β表达。敲除整合素β 1、Cdc 42和p110 β中的任何一种都抑制了PI 3 K信号传导的活性，并使DR-AsPC-1细胞对吉西他滨敏感。GST pull-down实验表明GTP-Cdc 42与p110 β相互作用。总之，这些数据表明，整合素β 1通过Cdc 42激活PI 3 K p110 β信号传导促进PDAC中的吉西他滨耐药性。体内实验也证实了这一结论。这些发现有助于更好地理解化疗耐药的分子机制，并促进更有针对性和有效的治疗策略的发展。(C)2018爱思唯尔公司All rights reserved.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most common malignancies with very poor prognosis due to its broad resistance to chemotherapy. Our previous study showed that integrin beta 1 expression is upregulated in PDAC and confers gemcitabine resistance in PDAC cells via the signaling pathway including Cdc42 and AKT activation. But the accurate signal transductions are not clear. Here, we aimed to illuminate the signal transductions of integrin 01 in the acquisition of gemcitabine resistance in PDAC. Drug-resistance (DR) cells from AsPC-1 parent cell line (PCL) were selected. Integrin beta 1 expression was determined using western blot assay. Changes in drug response and the activity of phosphatidylinositol 3-kinase (PI3K) signaling after knockdown of integrin beta 1, Cdc42 or p110 beta were evaluated using MTT, cleaved caspase-3 immunofluorescence and western blot assay. Western blot assays also detected the variations in Cdc42 activity and p110 beta expression after integrin 01 knockdown. The interaction between Cdc42 and p110 beta was determined by Glutathione S-transferase (GST) pull-down assay. The results showed that integrin beta 1 expression was upregulated in DR-AsPC-1 cells, and integrin beta 1 knockdown significantly decreased the activity of Cdc42, a target molecule of integrin beta 1, and p110 beta expression. Knockdown of anyone of integrin beta 1, Cdc42 and p110 beta inhibited the activity of PI3K signaling, and sensitized DR-AsPC-1 cells to gemcitabine. GST pull-down assay showed that GTP-Cdc42 interacted with p110 beta. Collectively, these data indicated that integrin beta 1 promoted gemcitabine resistance in PDAC through Cdc42 activation of PI3K p110 beta signaling. In vivo experiments also confirmed this conclusion. These findings contribute to a better understanding the molecular mechanism of chemoresistance and facilitate the development of more targeted and effective treatment strategy for PDAC. (C) 2018 Elsevier Inc. All rights reserved.