Loss of PDPK1 abrogates resistance to gemcitabine in label-retaining pancreatic cancer cells.
Loss of PDPK1 abrogates resistance to gemcitabine in label-retaining pancreatic cancer cells.
复制标题
PDPK1的损失消除了对标签胰腺癌细胞中吉西他滨的抵抗力。
DOI：
10.1186/s12885-018-4690-1
复制
发表时间：
2018-07-31
期刊：
影响因子：
3.8
通讯作者：
Rudloff U
中科院分区：
文献类型：
--
作者：
Li D;Mullinax JE;Aiken T;Xin H;Wiegand G;Anderson A;Thorgeirsson S;Avital I;Rudloff U
保留标记的癌细胞（LRCC）已被提议作为缓慢循环的癌症干细胞（CSC）的模型，其介导对化疗、肿瘤复发和转移的抵抗。迄今为止，LRCC 化疗耐药的分子机制仍不完全清楚。本研究旨在确定 LRCC 中的分子靶点，可用于克服对吉西他滨（一种治疗胰腺癌的标准化疗药物）的耐药性。通过流式细胞术对 Cy5-dUTP 染色后从胰腺癌细胞系中分离出 LRCC。从 LRCC、非 LRCC (NLRCC) 和大量肿瘤细胞获得的基因表达谱用于生成差异调节的通路网络。通过 RNAi 实验和药理学抑制评估 LRCC 中上调靶标的丢失对吉西他滨敏感性的影响。 PDPK1 是 LRCC 中上调的靶点之一，我们在患者的肿瘤样本中研究了 PDPK1 的表达模式，并将其与病理变量和临床结果相关。 LRCC 对吉西他滨的耐药性明显高于大量肿瘤细胞群。与非 LRCC 相比，非经典 EGF（表皮生长因子）介导的信号转导成为 LRCC 中最上调的网络，敲除 EGF 信号传导效应器 PDPK1（3-磷酸肌醇依赖性蛋白激酶 1）、BMX（BMX 非受体酪氨酸激酶）和 NTRK2（神经营养受体酪氨酸激酶 2）或用 PDPK1 抑制剂治疗可增加生长抑制并诱导细胞凋亡。吉西他滨。与非 LRCC 人群相比，PDPK1 的敲低优先增加吉西他滨治疗后的生长抑制并降低对诱导细胞凋亡的抵抗力。这些发现伴随着与手术切除标本中匹配的未受累胰腺相比，肿瘤中 PDPK1 的表达水平较低，并且 IHC 上的膜定位与高核级别呈负相关 (p<0.01)。与大块肿瘤细胞相比，胰腺癌细胞来源的 LRCC 对吉西他滨相对耐药，并且具有独特的转录组特征。 PDPK1 是 LRCC 中上调的 EGF 信号网络的成员之一，介导对吉西他滨的耐药性，被发现在胰腺癌标本中失调，可能是联合治疗研究中有吸引力的分子靶点。本文的在线版本 (10.1186/s12885-018-4690-1) 包含补充材料，可供授权用户使用。
Label-retaining cancer cells (LRCC) have been proposed as a model of slowly cycling cancer stem cells (CSC) which mediate resistance to chemotherapy, tumor recurrence, and metastasis. The molecular mechanisms of chemoresistance in LRCC remain to-date incompletely understood. This study aims to identify molecular targets in LRCC that can be exploited to overcome resistance to gemcitabine, a standard chemotherapy agent for the treatment of pancreas cancer. LRCC were isolated following Cy5-dUTP staining by flow cytometry from pancreatic cancer cell lines. Gene expression profiles obtained from LRCC, non-LRCC (NLRCC), and bulk tumor cells were used to generate differentially regulated pathway networks. Loss of upregulated targets in LRCC on gemcitabine sensitivity was assessed via RNAi experiments and pharmacological inhibition. Expression patterns of PDPK1, one of the upregulated targets in LRCC, was studied in patients’ tumor samples and correlated with pathological variables and clinical outcome. LRCC are significantly more resistant to gemcitabine than the bulk tumor cell population. Non-canonical EGF (epidermal growth factor)-mediated signal transduction emerged as the top upregulated network in LRCC compared to non-LRCC, and knock down of EGF signaling effectors PDPK1 (3-phosphoinositide dependent protein kinase-1), BMX (BMX non-receptor tyrosine kinase), and NTRK2 (neurotrophic receptor tyrosine kinase 2) or treatment with PDPK1 inhibitors increased growth inhibition and induction of apoptosis in response to gemcitabine. Knockdown of PDPK1 preferentially increased growth inhibition and reduced resistance to induction of apoptosis upon gemcitabine treatment in the LRCC vs non-LRCC population. These findings are accompanied by lower expression levels of PDPK1 in tumors compared to matched uninvolved pancreas in surgical resection specimens and a negative association of membranous localization on IHC with high nuclear grade (p < 0.01). Pancreatic cancer cell-derived LRCC are relatively resistant to gemcitabine and harbor a unique transcriptomic profile compared to bulk tumor cells. PDPK1, one of the members of an upregulated EGF-signaling network in LRCC, mediates resistance to gemcitabine, is found to be dysregulated in pancreas cancer specimens, and might be an attractive molecular target for combination therapy studies. The online version of this article (10.1186/s12885-018-4690-1) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
8
作者：
Jeanes, A.;Gottardi, C. J.;Yap, A. S.
通讯作者：
Yap, A. S.
影响因子：
7.4
作者：
Fillmore, Christine M.;Kuperwasser, Charlotte
通讯作者：
Kuperwasser, Charlotte
影响因子：
2.9
作者：
Mathews LA;Cabarcas SM;Hurt EM;Zhang X;Jaffee EM;Farrar WL
通讯作者：
Farrar WL
影响因子：
64.8
作者：
Fischer KR;Durrans A;Lee S;Sheng J;Li F;Wong ST;Choi H;El Rayes T;Ryu S;Troeger J;Schwabe RF;Vahdat LT;Altorki NK;Mittal V;Gao D
通讯作者：
Gao D
影响因子：
11.2
作者：
Li, Chenwei;Heidt, David G.;Simeone, Diane M.
通讯作者：
Simeone, Diane M.