HPV16 E6 enhances the radiosensitivity in HPV-positive human head and neck squamous cell carcinoma by regulating the miR-27a-3p/SMG1 axis.
HPV16 E6 enhances the radiosensitivity in HPV-positive human head and neck squamous cell carcinoma by regulating the miR-27a-3p/SMG1 axis.
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HPV16 E6通过调节miR-27a-3p/SMG1轴增强HPV阳性人头颈鳞状细胞癌的放射敏感性
DOI：
10.1186/s13027-021-00397-w
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发表时间：
2021-08-13
影响因子：
3.7
通讯作者：
Zhang C
中科院分区：
文献类型：
--
作者：
Long D;Xu L;Deng Z;Guo D;Zhang Y;Liu Z;Zhang C
头颈部鳞状细胞癌(HNSCC)是全球第六大常见的恶性肿瘤类型。人类乳头瘤病毒(HPV)阳性的HNSCC患者与HPV阴性的患者相比，放射敏感性显著增加。然而，HPV在HNSCC中的临床意义及其调控机制在很大程度上尚不清楚。本研究旨在探讨miR-27A-3p对HPV阳性HNSCC细胞放射敏感性的调控机制。方法建立E6高表达和E6基因敲除的HNSCC细胞株，用实时定量聚合酶链式反应(RT-qPCR)和免疫印迹法检测细胞的转染率。用RT-qPCR方法检测E6过表达载体或E6 siRNA对miR-27A-3p和DiGeorge综合征临界区8(Dgcr8)基因表达的影响。通过集落形成和TUNEL染色检测miR-27A-3p对HNSCC细胞放射敏感性的影响。利用生物信息学工具和荧光素酶报告基因分析鉴定SMG1是miR-27A-3p的直接靶点。结果HPVE6过表达上调HNSCC细胞中Dgcr8和miR-27A-3p的表达。接下来，我们证实了Dgcr8正向调控HNSCC细胞miR-27A-3p的表达。荧光素酶报告基因结果证实miR-27A-3P针对SMG1基因的3‘非编码区。MiR-27A-3p模拟载体转染组SMG1表达降低，miR-27A-3p抑制剂转染组SMG1表达增强。在miR-27A-3p模拟HNSCC细胞中，HNSCC细胞的凋亡活性明显高于对照HNSCC细胞。经4Gy射线照射后，转染miR-27A-3p抑制剂或SMG1高表达质粒的UM-SCC47细胞形成的克隆数明显多于相应的对照细胞。HPV16E6通过靶向miR-27A-3p/SMG1提高HNSCC细胞的放射敏感性。结论HPV16E6激活了Dgcr8/miR-27A-3p/SMG1轴，提高了HNSCC细胞的放射敏感性。我们的发现可能为提高HNSCC对放射治疗的反应提供一个新的治疗靶点。
BackgroundHead and neck squamous cell carcinoma (HNSCC) is the 6th most common malignant cancer type worldwide. Radiosensitivity has been shown to be significantly increased in patients with human papillomavirus (HPV)-positive HNSCC compared with HPV-negative patients. However, the clinical significance of HPV and its regulatory mechanisms in HNSCC are largely unknown. The aim of our study was to explore the regulatory mechanism of miR-27a-3p in the radiosensitivity of HPV-positive HNSCC cells.MethodsE6-overexpressing and E6-knockdown HNSCC cell lines were generated and the transfection efficiencies were evaluated by quantitative real-time PCR (RT-qPCR) and western blotting. The expression of miR-27a-3p and DiGeorge syndrome critical region 8 (DGCR8) was examined by RT-qPCR after transfection with E6 overexpressing plasmid or E6 siRNA. The effects of miR-27a-3p on the radiosensitivity of HNSCC cells were explored by a colony formation and TUNEL staining assays. Bioinformatic tools and luciferase reporter assays were used to identify that SMG1 is the direct target of miR-27a-3p. Furthermore, the effect of E6 overexpression on the regulation of the miR-27a-3p/SMG1 axis was investigated.ResultsIn our study, we found overexpression of HPV E6 upregulated the expression of DGCR8 and miR-27a-3p in HNSCC cells. We next confirmed that DGCR8 positively regulated the expression of miR-27a-3p in HNSCC cells. The luciferase reporter gene results verified that miR-27a-3p targeted the 3’UTR of SMG1 mRNA. MiR-27a-3p mimics transfection resulted in a decrease in SMG1 expression and miR-27a-3p inhibitor transfection increased SMG1 expression. Apoptotic activity of HNSCC cells was significantly increased in miR-27a-3p mimics HNSCC cells compared with control HNSCC cells. After treatment with 4 Gy irradiation, UM-SCC47 cells transfected with miR-27a-3p inhibitor or SMG1 overexpressing plasmid formed more colonies than the corresponding control cells. Furthermore, the rescue experiments demonstrated that HPV16 E6 improved the radiosensitivity of HNSCC cells by targeting miR-27a-3p/SMG1.ConclusionOur study demonstrated that HPV16 E6 activated the DGCR8/miR-27a-3p/SMG1 axis to enhance the radiosensitivity. Our findings might provide a novel therapeutic target to improve the response of HNSCC to radiotherapy.
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DOI：
10.26355/eurrev_202003_20523
发表时间：
2020-01-01
影响因子：
3.3
作者：
Cui, C-Y;Pan, Q-W;Ren, Z-X
通讯作者：
Ren, Z-X
影响因子：
3.5
作者：
Geng, Guangyong;Liu, Xin;Yuan, Xiao
通讯作者：
Yuan, Xiao
影响因子：
8
作者：
Pellom, Samuel T.;Rumfield, Claire Smalley;Jochems, Caroline
通讯作者：
Jochems, Caroline
影响因子：
5.6
作者：
Kiss F;Pohóczky K;Szállási A;Helyes Z
通讯作者：
Helyes Z
影响因子：
6.4
作者：
de Martel C;Plummer M;Vignat J;Franceschi S
通讯作者：
Franceschi S