Liu Tianyang, Ma Lin, Song Lianhao, Yan Bingqing, Zhang Siwei, Wang Bozhi, Zuo Nan, Sun Xinlei, Deng Yongqiang, Ren Qiushi, Li Yujun, Zhou Jingsong, Liu Qi, Wei Lanlan
Microbiology Department, Harbin Medical University, Harbin, Heilongjiang 150086, PR China; Shenzhen Bay Laboratory, Shenzhen, Guangdong 518132, PR China.
Department of Stomatology, Shenzhen University General Hospital, Shenzhen, Guangdong 518055, PR China.
Oral Oncol. 2022 Jun;129:105858. doi: 10.1016/j.oraloncology.2022.105858. Epub 2022 Apr 21.
This study aims to investigate how human papillomavirus (HPV) affects the key gene in the biological behaviors of head and neck squamous cell carcinoma (HNSCC) that leads to better response to radiotherapy.
The expression of key gene CENPM was analyzed using The Cancer Genome Atlas (TCGA) HNSCC data and HPV positive and HPV negative HNSCC tumors and cells. Assays with siRNAs, CRISPR/Cas9-based models, Western blot, qRT-PCR, ChIP, etc., were used to explore how HPV affects CENPM and response to radiotherapy for HNSCC.
CENPM occupies the hub in the HPV-related gene network. HPV-positive HNSCC showed higher level of CENPM expression comparing with HPV-negative HNSCC. HPV E5 has the most pronounced impact on CENPM (R = 0.44, p = 0.00081). This might result from the binding of transcription factor E2F1 to CENPM. We further found that inhibition of CENPM expression in HPV-positive HNSCC cell line SCC47 increased resistance to X-ray radiation by approximately 59% under 2 Gy irradiation, which may be resulted from a reduced proportion of mitotic cells.
HPV E5 enhances CENPM expression by transcription factor E2F1 in HNSCC, which results in a radiosensitive profile in cell cycle redistribution of HNSCC. Thus, HPV infection in HNSCC provides profound evidence that underscores the magnitude of E2F1 control of CENPM expression illustrating the potential clinical benefit of CENPM examination for difficult-to-treat HPV-negative cancers.
本研究旨在探讨人乳头瘤病毒（HPV）如何影响头颈部鳞状细胞癌（HNSCC）生物学行为中的关键基因，从而使其对放疗产生更好的反应。
利用癌症基因组图谱（TCGA）的HNSCC数据以及HPV阳性和HPV阴性的HNSCC肿瘤及细胞，分析关键基因CENPM的表达情况。采用小干扰RNA（siRNAs）、基于CRISPR/Cas9的模型、蛋白质免疫印迹法（Western blot）、实时定量聚合酶链反应（qRT-PCR）、染色质免疫沉淀法（ChIP）等实验方法，探究HPV如何影响CENPM以及HNSCC对放疗的反应。
CENPM在HPV相关基因网络中占据核心地位。与HPV阴性的HNSCC相比，HPV阳性的HNSCC中CENPM表达水平更高。HPV E5对CENPM的影响最为显著（R = 0.44，p = 0.00081）。这可能是由于转录因子E2F1与CENPM结合所致。我们进一步发现，在2 Gy照射下，抑制HPV阳性HNSCC细胞系SCC47中CENPM的表达可使细胞对X射线辐射的抗性增加约59%，这可能是由于有丝分裂细胞比例降低所致。
在HNSCC中，HPV E5通过转录因子E2F1增强CENPM的表达，从而在HNSCC细胞周期重分布中产生放射敏感表型。因此，HNSCC中的HPV感染充分证明了E2F1对CENPM表达控制的重要性，也说明了检测CENPM对难以治疗的HPV阴性癌症的潜在临床益处。
