Abstract
Biot2 is a novel murine testis-specific gene that was first identified using the SEREX technique, and named by our laboratory. Using conventional RT-PCR and real time RT-PCR, we tested the expression profile of Biot2 in normal tissues and various murine tumor cell lines. Using RNA interference, we studied the biological function of Biot2 in tumorigenesis. We applied various types of growth assay, such as the in vitro MTT, colony-forming and BrdU incorporation assays, along with in vivo tumorigenicity assays, to reveal its inhibition of tumor cell proliferation. The results revealed that the Biot2 transcript was detected only and strongly in the testis tissues and abundantly in five types of murine cancer cell line. Treating B16 murine melanoma, LL/2 murine Lewis lung carcinoma and CT26 murine colorectal adenocarcinoma with special shRNA targeting Biot2 can significantly reduce the proliferation rate of these three tumor cell lines in vitro, as measured by the MTT, colony-forming and BrdU incorporation assays. The tumorigenicity of the CT26 cells transfected with special shRNA targeting Biot2 was also decreased distinctly in vivo compared with the control. It was therefore concluded that Biot2 plays a key role in tumorigenesis and could be a potential target for biotherapy.
Key words: RNA interference, Biot2, Testis-specific, Proliferation
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Abbreviations used
- ATCC
American Type Culture Collection
- BrdU
bromodeoxyuridine
- B16
murine melanoma cell
- CT antigen
cancer/testis antigen
- CT26
murine colorectal adenocarcinoma cell
- Hepa and H22
murine hepatocellular carcinoma cell
- LL/2
murine Lewis lung carcinoma cell
- MethA
murine fibrosarcoma cell
- NCBI
National Center for Biotechnology Information
- OD
optical density
- ORF
open reading frame
- SD
standard diviation
- SF
survival fraction
- shRNA
short hairpin RNA
- 4T1
murine breast cancer cells
Footnotes
These authors contributed equally to this research
Contributor Information
Feng Peng, Phone: +86 28 85164063, FAX: +86 28 85164060, Email: evenforeven@yahoo.com.cn.
Han-Shuo Yang, Phone: +86 28 85164063, FAX: +86 28 85164060, Email: yhansh@scu.edu.cn.
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