Dataset Information

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

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 in vitro. Moreover, it could also inhibit the growth of xenografts and increase the apoptosis index and radiosensitivity in vivo. 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 in vitro and in vivo.

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 in vitro 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 in vitro and in vivo.

SUBMITTER: Chen K

PROVIDER: S-EPMC7803545 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

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

Chen Kaihua K Li Ling L Qu Song S Pan Xinbin X Sun Yongchu Y Wan Fangzhu F Yu Binbin B Zhou Lei L Zhu Xiaodong X

Aging 20201124 24

<h4>Objective</h4>This study aimed to explore the effect of silencing hTERT on the CSC-like characteristics and radioresistance of CNE-2R cells.<h4>Results</h4>Silencing hTERT suppressed CNE-2R cell proliferation and increased the cell apoptosis rate and radiosensitivity <i>in vitro</i>. Moreover, it could also inhibit the growth of xenografts and increase the apoptosis index and radiosensitivity <i>in vivo</i>. Further study discovered that after silencing hTERT, telomerase activity in CNE-2R c ...[more]

PMID: 33234740

Similar Datasets

Project description:Nasopharyngeal carcinoma (NPC) is a common malignant tumor. In recent studies, we demonstrated that overexpression of the Bax inhibitor-1 (BI-1) induces cell transformation in NIH3T3 cells and that knockdown of BI-1 and human telomerase reverse transcriptase (hTERT) gene expression suppresses NPC cell proliferation and induces apoptosis. To evaluate the combination anti-tumor effects of siRNAs against hTERT and BI-1 in the CNE-2 NPC cell line, combined and separate short-hairpin (sh)RNA plasmids targeting hTERT and BI-1, respectively, were constructed. hTERT and BI-1 mRNA and protein levels were examined by real-time polymerase chain reaction (PCR) and western blot analysis. Cell proliferation, colony formation and migration ability were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), soft agar and wound healing assay. Cell apoptosis was observed by flow cytometry, Hoechst 33258 staining and caspase-3 activity. hTERT, BI-1 and combined shRNA plasmids were injected into xenograft NPC tumor tissues, and expression of hTERT and BI-1 was detected by real-time PCR and immunohistochemistry. Tumor growth was measured by tumor volume and apoptosis in vivo was confirmed by TdT-mediated dUTP nick end-labeling (TUNEL). Our results showed that combined shRNA specific for hTERT and BI-1 markedly suppressed hTERT and BI-1 gene expression in vitro and in vivo. In addition, CNE-2 cell proliferation was inhibited in vitro as well as in vivo. Following the knockdown of the two gene expressions, CNE-2 exhibited a decrease in colony formation and migration ability and an increase in the apoptotic rate compared to the control groups. Our in vitro and in vivo study showed that the combinative silencing of the two genes enhanced the therapeutic effect compared to the silencing of each individual shRNA. These data suggested that combinatorial gene therapy targeting hTERT and BI-1 may be beneficial as a tumor therapy strategy against human NPCs.

Dataset Information

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

Objective

Results

Conclusion

Methods

Publications

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

Similar Datasets

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