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沉默端粒酶逆转录酶可减弱耐辐射鼻咽癌细胞系 CNE-2R 的肿瘤干细胞样特征和辐射抗性。

Silencing hTERT attenuates cancer stem cell-like characteristics and radioresistance in the radioresistant nasopharyngeal carcinoma cell line CNE-2R.

机构信息

Department of Radiation Oncology, Guangxi Medical University Cancer Hospital, Nanning 530021, Guangxi, China.

Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor (Guangxi Medical University), Ministry of Education, Nanning 530021, Guangxi, China.

出版信息

Aging (Albany NY). 2020 Nov 24;12(24):25599-25613. doi: 10.18632/aging.104167.

Abstract

OBJECTIVE

This study aimed to explore the effect of silencing hTERT on the CSC-like characteristics and radioresistance of CNE-2R cells.

RESULTS

Silencing hTERT suppressed CNE-2R cell proliferation and increased the cell apoptosis rate and radiosensitivity . Moreover, it could also inhibit the growth of xenografts and increase the apoptosis index and radiosensitivity . Further study discovered that after silencing hTERT, telomerase activity in CNE-2R cells was markedly suppressed, along with remarkably down-regulated stem cell-related protein levels both and .

CONCLUSION

Silencing hTERT can suppress the CSC-like characteristics of CNE-2R cells to enhance their radiosensitivity, revealing that hTERT may become a potential target for treating radioresistant NPC.

METHODS

An RNAi lentiviral vector specific to the hTERT gene was constructed to infect CNE-2R cells, the hTERT silencing effect was verified through qPCR and Western blot assays, and telomerase activity was detected by PCR-ELISA. Moreover, radiosensitivity was detected through colony formation assays, CCK-8 assays and flow cytometry. Tumor growth and radioresistance were also evaluated using xenograft models, while the apoptosis index in xenografts was measured through TUNEL assay. Levels of stem cell-related proteins were determined and .

摘要

目的

本研究旨在探讨沉默端粒酶逆转录酶(hTERT)对鼻咽癌(NPC)CNE-2R 细胞癌干细胞(CSC)样特性和放射抵抗性的影响。

结果

沉默 hTERT 可抑制 CNE-2R 细胞增殖,增加细胞凋亡率和放射敏感性。此外,还能抑制异种移植瘤的生长,增加凋亡指数和放射敏感性。进一步研究发现,沉默 hTERT 后,CNE-2R 细胞中端粒酶活性明显受到抑制,同时干细胞相关蛋白水平也显著下调。

结论

沉默 hTERT 可抑制 CNE-2R 细胞的 CSC 样特性,增强其放射敏感性,提示 hTERT 可能成为治疗 NPC 放射抵抗的潜在靶点。

方法

构建靶向 hTERT 基因的 RNAi 慢病毒载体感染 CNE-2R 细胞,通过 qPCR 和 Western blot 验证 hTERT 沉默效果,PCR-ELISA 检测端粒酶活性。此外,通过集落形成实验、CCK-8 实验和流式细胞术检测放射敏感性。利用异种移植模型评估肿瘤生长和放射抵抗性,通过 TUNEL 实验检测异种移植瘤中的凋亡指数。通过 Western blot 检测干细胞相关蛋白水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/38a7/7803545/8f720c5451c1/aging-12-104167-g001.jpg

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