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苯并[a]芘对口腔鳞状细胞癌细胞增殖和转移的影响:基于RNA测序的转录组分析

Effect of benzo[a]pyrene on proliferation and metastasis of oral squamous cell carcinoma cells: A transcriptome analysis based on RNA-seq.

作者信息

He Hanyi, Huang Yixing, Lu Yueyue, Wang Xinlu, Ni Haifeng, Wu Yihua, Xia Dajing, Ye Dong, Ding Jinwang, Mao Yanjiao, Teng Yaoshu

机构信息

Department of Otorhinolaryngology, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, China.

The Fourth Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, China.

出版信息

Environ Toxicol. 2022 Nov;37(11):2589-2604. doi: 10.1002/tox.23621. Epub 2022 Jul 23.

Abstract

Benzo[a]pyrene (BaP), a representative polycyclic aromatic hydrocarbon compound, is a carcinogen that causes head and neck cancers. Despite intensive research, the molecular mechanism of BaP in the development of oral squamous cell carcinoma (OSCC) remains largely unknown. In the present study, the SCC-9 human OSCC cell line was cultured in vitro, separated into treatment groups, and treated with dimethyl sulfoxide or BaP at various concentrations. The malignant behavior ascribed to the BaP treatment was investigated by cell proliferation, clony formation assay, and Transwell assays. Furthermore, transcriptome sequencing was performed to detect the differentially expressed genes, followed by quantitative real-time PCR to measure the expression levels of nine of these genes. Moreover, the Gene Ontology (GO) term and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed the biological processes and signaling pathways in which the target genes were involved. Significant effects on SCC-9 cell proliferation, tumorigenicity, cell migration, and invasion were observed after exposure to 8 μM BaP. Additional results revealed that BaP inhibited apoptosis in a dose-dependent manner. The transcriptome sequencing results showed 137 upregulated genes and 135 downregulated genes induced by BaP, associated with tumor-related biological processes and signaling pathways, mainly including transcriptional dysregulation in cancer, the tumor necrosis factor signaling pathway, metabolism of xenobiotics by cytochrome P450, mitogen-activated protein kinase signaling pathway, and so forth. Our study demonstrates that BaP may regulate the expression of certain genes involved in tumor-associated signaling pathways, thereby promoting the proliferative, tumorigenic, and metastatic behaviors of OSCC cells while suppressing their apoptosis.

摘要

苯并[a]芘(BaP)是一种具有代表性的多环芳烃化合物,是一种可引发头颈癌的致癌物。尽管进行了深入研究,但BaP在口腔鳞状细胞癌(OSCC)发生发展中的分子机制仍 largely未知。在本研究中,SCC-9人OSCC细胞系在体外培养,分为处理组,并用二甲基亚砜或不同浓度的BaP处理。通过细胞增殖、克隆形成试验和Transwell试验研究了BaP处理所致的恶性行为。此外,进行转录组测序以检测差异表达基因,随后进行定量实时PCR以测量其中9个基因的表达水平。此外,基因本体(GO)术语和京都基因与基因组百科全书(KEGG)通路分析显示了靶基因所涉及的生物学过程和信号通路。暴露于8 μM BaP后,观察到对SCC-9细胞增殖、致瘤性、细胞迁移和侵袭有显著影响。其他结果显示,BaP以剂量依赖性方式抑制细胞凋亡。转录组测序结果显示,BaP诱导137个上调基因和135个下调基因,这些基因与肿瘤相关生物学过程和信号通路有关,主要包括癌症中的转录失调、肿瘤坏死因子信号通路、细胞色素P450对外源生物的代谢、丝裂原活化蛋白激酶信号通路等。我们的研究表明,BaP可能调节某些参与肿瘤相关信号通路的基因的表达,从而促进OSCC细胞的增殖、致瘤和转移行为,同时抑制其凋亡。

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