Cigarette Smoke Induces Stem Cell Features of Pancreatic Cancer Cells via PAF1.
Cigarette Smoke Induces Stem Cell Features of Pancreatic Cancer Cells via PAF1.
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DOI：
10.1053/j.gastro.2018.05.041
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发表时间：
2018-09
期刊：
影响因子：
29.4
通讯作者：
Ponnusamy MP
中科院分区：
文献类型：
--
作者：
Nimmakayala RK;Seshacharyulu P;Lakshmanan I;Rachagani S;Chugh S;Karmakar S;Rauth S;Vengoji R;Atri P;Talmon GA;Lele SM;Smith LM;Thapa I;Bastola D;Ouellette MM;Batra SK;Ponnusamy MP
吸烟是胰腺癌的主要危险因素。侵袭性胰腺肿瘤含有具有干细胞特征的癌细胞。我们研究了香烟烟雾是否会诱导胰腺癌细胞的干细胞特征。KrasG12D;Pdx1-Cre （KC）小鼠暴露于香烟烟雾或清洁空气（对照组）长达20周；收集胰腺标本，进行组织学、定量反转录PCR和共聚焦免疫荧光显微镜分析。将HPNE和Capan1细胞暴露于香烟烟雾提取物（CSE）、尼古丁和尼古丁衍生致癌物（NNN或NNK）或清洁空气（对照组）中80天，并使用基于流式细胞术的自身荧光、球形成和免疫印迹法评估干细胞标记物和特征。细胞中的蛋白质被小干扰rna敲低。我们对暴露于cse的细胞进行RNA测序分析。我们用染色质免疫沉淀法证实了FOS like 1，ap -1转录因子亚基（FOSL1）与RNA聚合酶ii相关因子（PAF1）启动子的结合。我们获得胰腺导管腺癌（PDAC）和匹配的非肿瘤组织（n=15），并进行免疫组织化学分析。与对照细胞相比，HPNE和Capan1细胞长期暴露于CSE中导致其干细胞标记物增加，包括自身荧光和球体形成。这些细胞通过胆碱能受体烟碱α 7亚基（CHRNA7）信号传导至丝裂原活化蛋白激酶1和FOSL1，从而增加ABCG2、SOX9和PAF1的表达。PAF1基因敲低的胰腺细胞系在暴露于CSE后没有出现干细胞的特征。细胞暴露于NNN和NNK后，CHRNA7、FOSL1和PAF1的表达随着干细胞特征的增加而增加。与对照小鼠相比，暴露于香烟烟雾的KC小鼠胰腺中PAF1 mRNA和蛋白水平升高，SOX9的表达也增加。与非肿瘤胰腺组织相比，PDAC组织中PAF1和FOSL1水平升高，尤其是来自吸烟者的组织。CSE暴露增加了多能转录因子PHD指蛋白5A的表达及其与PAF1的相互作用。暴露于香烟烟雾通过CHRNA7信号和FOSL1激活PAF1表达激活胰腺细胞的干细胞特征。长期暴露于香烟烟雾的人类和小鼠胰腺肿瘤中，PAF1水平升高。
Cigarette smoking is a major risk factor for pancreatic cancer. Aggressive pancreatic tumors contain cancer cells with stem cell features. We investigated whether cigarette smoke induces stem cell features in pancreatic cancer cells. KrasG12D; Pdx1-Cre (KC) mice were exposed to cigarette smoke or clean air (controls) for up to 20 weeks; pancreata were collected and analyzed by histology, quantitative reverse transcription PCR, and confocal immunofluorescence microscopy. HPNE and Capan1 cells were exposed to cigarette smoke extract (CSE), nicotine and nicotine-derived carcinogens (NNN or NNK), or clean air (controls) for 80 days and evaluated for stem cell markers and features using flow cytometry-based autofluorescence, sphere formation, and immunoblot assays. Proteins were knocked down in cells with small interfering RNAs. We performed RNA sequencing analyses of CSE-exposed cells. We used chromatin immunoprecipitation assays to confirm the binding of FOS like 1, AP-1 transcription factor subunit (FOSL1) to RNA polymerase II-associated factor (PAF1) promoter. We obtained pancreatic ductal adenocarcinoma (PDAC) and matched non-tumor tissues (n=15) and performed immunohistochemical analyses. Chronic exposure of HPNE and Capan1 cells to CSE caused them to increase markers of stem cells, including autofluorescence and sphere formation, compared to control cells. These cells increased expression of ABCG2, SOX9 and PAF1, via cholinergic receptor nicotinic alpha 7 subunit (CHRNA7) signaling to mitogen-activated protein kinase 1 and FOSL1. Pancreatic cell lines with knockdown of PAF1 did not develop features of stem cells upon exposure to CSE. Exposure of cells to NNN and NNK led to increased expression of CHRNA7, FOSL1, and PAF1 along with stem cell features. Pancreata from KC mice exposed to cigarette smoke had increased levels of PAF1 mRNA and protein, compared with control mice, as well as increased expression of SOX9. Levels of PAF1 and FOSL1 were increased in PDAC tissues, especially those from smokers, compared with non-tumor pancreatic tissue. CSE exposure increased expression of PHD finger protein 5A, a pluripotent transcription factor and its interaction with PAF1. Exposure to cigarette smoke activates stem cell features of pancreatic cells, via CHRNA7 signaling and FOSL1 activation of PAF1 expression. Levels of PAF1 are increased in pancreatic tumors of humans and mice with chronic cigarette smoke exposure.
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影响因子：
4.3
作者：
Little MP;Hendry JH
通讯作者：
Hendry JH
DOI：
10.5114/wo.2016.63043
发表时间：
2017
期刊：
Contemporary oncology (Poznan, Poland)
影响因子：
--
作者：
Kuroczycki-Saniutycz S;Grzeszczuk A;Zwierz ZW;Kołodziejczyk P;Szczesiul J;Zalewska-Szajda B;Ościłowicz K;Waszkiewicz N;Zwierz K;Szajda SD
通讯作者：
Szajda SD
影响因子：
8
作者：
Moniaux, N.;Nemos, C.;Batra, S. K.
通讯作者：
Batra, S. K.
影响因子：
16
作者：
Penheiter, KL;Washburn, TM;Jaehning, JA
通讯作者：
Jaehning, JA
影响因子：
16
作者：
Mueller, CL;Porter, SE;Jaehning, JA
通讯作者：
Jaehning, JA