ZIP4 Increases Expression of Transcription Factor ZEB1 to Promote Integrin α3β1 Signaling and Inhibit Expression of the Gemcitabine Transporter ENT1 in Pancreatic Cancer Cells.
ZIP4 Increases Expression of Transcription Factor ZEB1 to Promote Integrin α3β1 Signaling and Inhibit Expression of the Gemcitabine Transporter ENT1 in Pancreatic Cancer Cells.
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DOI：
10.1053/j.gastro.2019.10.038
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发表时间：
2020-02
期刊：
影响因子：
29.4
通讯作者：
Li M
中科院分区：
文献类型：
--
作者：
Liu M;Zhang Y;Yang J;Cui X;Zhou Z;Zhan H;Ding K;Tian X;Yang Z;Fung KA;Edil BH;Postier RG;Bronze MS;Fernandez-Zapico ME;Stemmler MP;Brabletz T;Li YP;Houchen CW;Li M
胰腺肿瘤经历快速生长和进展，变得对化疗耐药，并且在手术后复发。我们研究了溶质载体家族39成员4（SLC 39 A4，也称为ZIP 4）的功能，该家族调节细胞内锌的浓度，并在胰腺癌细胞、细胞系和小鼠中增加。我们从接受手术和吉西他滨化疗的患者中获得了93份胰腺癌标本（肿瘤和邻近的非肿瘤组织），并通过免疫组织化学对其进行了分析。在AsPC-1和MIA PaCa-2胰腺癌细胞系以及KPC和KPC-ZEB 1敲除小鼠的胰腺细胞中，ZIP 4和/或ITGA 3或ITGB 1过表达或用小发夹RNA敲低，并建立胰腺球状体;通过免疫印迹、逆转录PCR和液相色谱串联质谱法分析细胞和球状体。我们使用染色质沉淀和荧光素酶报告基因分析研究了ZIP 4对ZEB 1、ITGA 3、ITGB 1、JNK和ENT 1的转录调控。向裸鼠注射遗传操作的AsPC-1和MIA PaCa-2细胞，并测量异种移植肿瘤和转移的生长。在来自患者的胰腺癌标本中，ZIP 4水平的增加与生存时间的缩短相关。过表达ZIP 4的MIA PaCa-2细胞对吉西他滨、5-FU和顺铂的耐药性增加，而ZIP 4敲低的AsPC-1细胞对这些药物的敏感性增加。在小鼠中，从ZIP 4敲低的AsPC-1细胞生长的异种移植肿瘤更小，对吉西他滨更敏感。ZIP 4过表达显著降低了吉西他滨在胰腺癌细胞中的积累，增加了小鼠异种移植肿瘤的生长，并增加了整合素亚基ITGA 3和ITGB 1的表达;在ZIP 4敲低的细胞中，ITGA 3和ITGB 1的表达降低。尽管ZIP 4过表达，但ITGA 3或ITGB 1敲减的胰腺癌细胞在小鼠中增殖减少并形成较小的肿瘤;从这些细胞建立的球状体对吉西他滨的敏感性增加。我们发现ZIP 4激活STAT 3以诱导ZEB 1的表达，ZEB 1诱导KPC细胞中ITGA 3和ITGB 1的表达。ITGA 3和ITGB 1表达增加以及随后通过JNK的整合素α3β1信号传导抑制吉西他滨转运蛋白ENT 1的表达，从而减少胰腺癌细胞对吉西他滨的摄取。ZEB 1敲除细胞对吉西他滨的敏感性增加。在胰腺癌细胞系和小鼠的研究中，我们发现ZIP 4增加了转录因子ZEB 1的表达，ZEB 1激活了ITGA 3和ITGB 1的表达。随后通过JNK增加整合素α3β1信号传导，抑制吉西他滨转运蛋白ENT 1的表达，从而使细胞摄取更少量的药物。该通路的激活可能有助于介导胰腺肿瘤对化疗药物的耐药性。我们鉴定了在胰腺癌细胞中激活的信号通路，其介导了它们对化疗剂如吉西他滨的抗性。
Pancreatic tumors undergo rapid growth and progression, become resistant to chemotherapy, and recur after surgery. We studied the functions of the solute carrier family 39 member 4 (SLC39A4, also called ZIP4), which regulates concentrations of intracellular zinc and is increased in pancreatic cancer cells, in cell lines and mice. We obtained 93 pancreatic cancer specimens (tumor and adjacent non-tumor tissues) from patients who underwent surgery and gemcitabine chemotherapy and analyzed them by immunohistochemistry. ZIP4 and/or ITGA3 or ITGB1 were overexpressed or knocked down with small hairpin RNAs in AsPC-1 and MIA PaCa-2 pancreatic cancer cells lines, and in pancreatic cells from KPC and KPC-ZEB1 knockout mice, and pancreatic spheroids were established; cells and spheroids were analyzed by immunoblots, reverse transcription PCR, and liquid chromatography tandem mass spectrometry. We studied transcriptional regulation of ZEB1, ITGA3, ITGB1, JNK, and ENT1 by ZIP4 using chromatin precipitation and luciferase reporter assays. Nude mice were given injections of genetically manipulated AsPC-1 and MIA PaCa-2 cells and growth of xenograft tumors and metastases was measured. In pancreatic cancer specimens from patients, increased levels of ZIP4 associated with shorter survival times. MIA PaCa-2 cells that overexpressed ZIP4 had increased resistance to gemcitabine, 5-FU, and cisplatin, whereas AsPC-1 cells with ZIP4 knockdown had increased sensitivity to these drugs. In mice, xenograft tumors grown from AsPC-1 cells with ZIP4 knockdown were smaller and more sensitive to gemcitabine. ZIP4 overexpression significantly reduced accumulation of gemcitabine in pancreatic cancer cells, increased growth of xenograft tumors in mice, and increased expression of the integrin subunits ITGA3 and ITGB1; expression of ITGA3 and ITGB1 was reduced in cells with ZIP4 knockdown. Pancreatic cancer cells with ITGA3 or ITGB1 knockdown had reduced proliferation and formed smaller tumors in mice, despite overexpression of ZIP4; spheroids established from these cells had increased sensitivity to gemcitabine. We found ZIP4 to activate STAT3 to induce expression of ZEB1, which induced expression of ITGA3 and ITGB1 in KPC cells. Increased ITGA3 and ITGB1 expression and subsequent integrin α3β1 signaling, via JNK, inhibited expression of the gemcitabine transporter ENT1, which reduced gemcitabine uptake by pancreatic cancer cells. ZEB1-knockdown cells had increased sensitivity to gemcitabine. In studies of pancreatic cancer cell lines and mice, we found that ZIP4 increases expression of the transcription factor ZEB1, which activates expression of ITGA3 and ITGB1. The subsequent increase in integrin α3β1 signaling, via JNK, inhibits expression of the gemcitabine transporter ENT1, so that cells take up smaller amounts of the drug. Activation of this pathway might help mediate resistance of pancreatic tumors to chemotherapeutic agents. We identified a signaling pathway that is activated in pancreatic cancer cells that mediates their resistance to chemotherapeutic agents such as gemcitabine.
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DOI：
10.1073/pnas.0709307104
发表时间：
2007-11-20
影响因子：
11.1
作者：
Li, Min;Zhang, Yuqing;Yao, Qizhi
通讯作者：
Yao, Qizhi
影响因子：
37.3
作者：
Lin L;Deangelis S;Foust E;Fuchs J;Li C;Li PK;Schwartz EB;Lesinski GB;Benson D;Lü J;Hoyt D;Lin J
通讯作者：
Lin J
DOI：
10.1155/2012/283181
发表时间：
2012
期刊：
Chemotherapy research and practice
影响因子：
--
作者：
Aoudjit F;Vuori K
通讯作者：
Vuori K
影响因子：
4.8
作者：
Hagmann, Wolfgang;Jesnowski, Ralf;Loehr, Johannes Matthias
通讯作者：
Loehr, Johannes Matthias
影响因子：
15.9
作者：
Koay, Eugene J.;Truty, Mark J.;Fleming, Jason B.
通讯作者：
Fleming, Jason B.