Construction of a tumor‑specific bioluminescent eukaryotic expression vector and analysis of its expression 
in vitro and in vivo

Chen,Qian; Li,Fen; Zhou,Fuxiang; Wang,Wenbo; Xu,Yu; Sun,Wenjie; Zhou,Yunfeng

doi:10.3892/ol.2013.1343

July 2013 Volume 6 Issue 1

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July 2013 Volume 6 Issue 1

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Article

Construction of a tumor‑specific bioluminescent eukaryotic expression vector and analysis of its expression in vitro and in vivo

Authors:
- Qian Chen
- Fen Li
- Fuxiang Zhou
- Wenbo Wang
- Yu Xu
- Wenjie Sun
- Yunfeng Zhou
View Affiliations / Copyright

Affiliations: Department of Radiation and Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center and Key Laboratory of Tumor Biological Behavior, Wuhan, Hubei 430071, P.R. China
Pages: 207-211
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Published online on: May 8, 2013

https://doi.org/10.3892/ol.2013.1343
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Abstract

The aims of this study were to construct a tumor‑specific bioluminescent eukaryotic vector driven by the hTERT gene promoter and to establish a stable HeLa cell line expressing a modified firefly luciferase gene. PhTERTp‑luc and pGL4.17 (luc2/Neo) were digested with SacI and HindIII, respectively, and the recombinant vector phTERTp‑luc‑neo was generated by ligating the desired fragments. The expression of phTERTp‑luc‑neo was tested in a non‑transformed cell line (MRC‑5), and in telomerase‑positive (HeLa, MCF‑7 and 293T) and ‑negative (U2OS and SaOS) transformed cell lines using a luciferase assay. Results showed that the recombinant vector had higher luciferase activity in telomerase‑positive transformed cell lines. PhTERTp‑luc‑neo was transfected into a HeLa cell line, selected by G418 and bioluminescence imaging, and a cell clone HeLa‑luc that constitutively expressed both neomycin and luciferase was obtained. We also conducted experiments in animals to observe luciferase activity in vivo using stable cell lines that were subcutaneously implanted into BALB/c nude mice and tumor growth was monitored by bioluminescence imaging. The HeLa‑luc cell line retained its oncogenicity and tumors were detected on the fifth day following implantation by bioluminescence imaging. This study has formed a basis for the study of the expression and regulation of hTERT and early tumor detection. It also provides a convenient, sensitive and reliable platform for cervical cancer research.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Construction of a tumor‑specific bioluminescent eukaryotic expression vector and analysis of its expression in vitro and in vivo

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