Tobacco Exposure Enhances Human Papillomavirus 16 Oncogene Expression via EGFR/PI3K/Akt/c-Jun Signaling Pathway in Cervical Cancer Cells
Tobacco Exposure Enhances Human Papillomavirus 16 Oncogene Expression via EGFR/PI3K/Akt/c-Jun Signaling Pathway in Cervical Cancer Cells
复制标题
烟草暴露通过EGFR/PI3K/Akt/c-jun信号通路促进宫颈癌细胞中HPV16癌基因的表达
DOI：
10.3389/fmicb.2018.03022
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发表时间：
2018-12-17
影响因子：
5.2
通讯作者：
Aguayo, Francisco
中科院分区：
文献类型：
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作者：
Munoz, Juan P.;Carrillo-Beltran, Diego;Aguayo, Francisco
高危型人乳头瘤病毒（HR-HPV）感染不是宫颈癌发展的充分条件，因为大多数感染是良性的，自然清除。流行病学研究表明，吸烟是宫颈癌发生和进展的HR-HPV的辅助因素，尽管烟草烟雾与HR-HPV在这种恶性肿瘤中的合作机制尚不清楚。由于HR-HPV E6/E7癌蛋白在宫颈癌中过表达与香烟烟雾成分（CSC）共定位，在这项研究中，我们解决了参与两种致癌物质之间潜在相互作用的信号通路。宫颈癌衍生的细胞系CaSki（HPV 16;每个细胞500个拷贝）和SiHa（HPV 16;每个细胞2个拷贝）急性暴露于各种无毒浓度的CSC，我们发现E6和E7水平以剂量依赖性方式显著增加。使用一个报告构建体，包含荧光素酶基因的控制下的完整的HPV 16长控制区（LCR），我们还发现，p97启动子活性是依赖于CSC。LCR-驻留TPA（12-O-十四烷酰基佛波醇13-乙酸酯）-响应元件（TRE）中的非同义突变显著降低了p97启动子激活。磷酸化蛋白质组学阵列和特异性抑制剂显示CSC介导的E6/E7过表达至少部分依赖于EGFR磷酸化。此外，我们发现PI 3 K/Akt通路对于CSC诱导的E6/E7过表达至关重要。最后，我们证明了HPV 16 E6/E7过表达是由JUN过表达、c-Jun磷酸化和该转录因子募集到HPV 16 LCR中的TRE位点介导的。我们的结论是，急性暴露于烟草烟雾激活HPV 16 E6和E7癌基因的转录通过p97启动子激活，这涉及EGFR/PI 3 K/Akt/C-Jun信号通路激活在宫颈癌细胞。
High-risk human papillomavirus (HR-HPV) infection is not a sufficient condition for cervical cancer development because most infections are benign and naturally cleared. Epidemiological studies revealed that tobacco smoking is a cofactor with HR-HPV for cervical cancer initiation and progression, even though the mechanism by which tobacco smoke cooperates with HR-HPV in this malignancy is poorly understood. As HR-HPV E6/E7 oncoproteins overexpressed in cervical carcinomas colocalize with cigarette smoke components (CSC), in this study we addressed the signaling pathways involved in a potential interaction between both carcinogenic agents. Cervical cancer-derived cell lines, CaSki (HPV16; 500 copies per cell) and SiHa (HPV16; 2 copies per cell), were acutely exposed to CSC at various non-toxic concentrations and we found that E6 and E7 levels were significantly increased in a dose-dependent manner. Using a reporter construct containing the luciferase gene under the control of the full HPV16 long control region (LCR), we also found that p97 promoter activity is dependent on CSC. Non-synonymous mutations in the LCR-resident TPA (12-O-tetradecanoylphorbol 13-acetate)-response elements (TRE) had significantly decreased p97 promoter activation. Phosphoproteomic arrays and specific inhibitors revealed that CSC-mediated E6/E7 overexpression is at least in part reliant on EGFR phosphorylation. In addition, we showed that the PI3K/Akt pathway is crucial for CSC-induced E6/E7 overexpression. Finally, we demonstrated that HPV16 E6/E7 overexpression is mediated by JUN. overexpression, c-Jun phosphorylation and recruitment of this transcription factor to TRE sites in the HPV16 LCR. We conclude that acute exposure to tobacco smoke activates the transcription of HPV16 E6 and E7 oncogenes through p97 promoter activation, which involves the EGFR/PI3K/Akt/C-Jun signaling pathway activation in cervical cancer cells.