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RNA interference against Biot2, a novel mouse testis — specific gene, inhibits the growth of tumor cells

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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.

Abbreviations

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

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Author information

Correspondence to Feng Peng or Han-Shuo Yang.

Additional information

These authors contributed equally to this research

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Key words

  • RNA interference
  • Biot2
  • Testis-specific
  • Proliferation