Knock-out transmembrane prostate androgen-induced protein gene suppressed triple-negative breast cancer cell proliferation

  • Bantari W.K. Wardhani Doctoral Program in Biomedicine, Faculty of Medicine, Universitas Indonesia, Jakarta
  • Meidi U. Puteri Medical Science Master Program, Graduate School of Comprehensive Human Science, University of Tsukuba
  • Yukihide Watanabe Departement of Experimental Pathology, Faculty of Medicine, University of Tsukuba
  • Melva Louisa Departement of Pharmacology and Therapeutics, Faculty of Medicine, Universitas Indonesia, Jakarta
  • Rianto Setiabudy Departement of Pharmacology and Therapeutics, Faculty of Medicine, Universitas Indonesia, Jakarta
  • Mitsuyasu Kato Departement of Experimental Pathology, Faculty of Medicine, University of Tsukuba
Keywords: proliferation, TGF-β, TMEPAI, triple negative breast cancer
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Abstract

Background: Triple negative breast cancer (TNBC) tends to grow more rapidly and has poorer prognosis compared to others. High expression of transmembrane prostate androgen-induced protein (TMEPAI) correlates with poor prognosis in TNBC patients. However, the mechanistic role of TMEPAI in tumorigenic remains unknown. This study aimed to knock-out TMEPAI in TNBC cell line to determine its function further in cells proliferation.

Methods: CRISPR-Cas9 has been used previously to knock-out TMEPAI in Hs857T TNBC cell line. Hs587T TNBC parental cell line (wild-type/WT) and TMEPAI knock out Hs 586T cell lines were cultured in Dulbecco’s modified eagle medium (DMEM) supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin and amphotericin B. Both cell lines were seeded in 24-well plates and counted every two days, then proliferation rates were plotted. Afterwards, total RNA were isolated from the cells and Ki-67, and TGF-β mRNA expression levels as proliferation markers were determined.

Results: Cell proliferation rates as displayed in growth curve plots showed that WT-TMEPAI cell line grew more rapidly than KO-TMEPAI. In accordance, mRNA expression levels of  Ki-67 and TGF-β  were significantly decreased KO-TMEPAI as compare to TMEPAI-WT.

Conclusion: Knock-out of TMEPAI attenuates cell proliferation in TNBC.

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Published
2017-11-27
How to Cite
1.
Wardhani BW, Puteri MU, Watanabe Y, Louisa M, Setiabudy R, Kato M. Knock-out transmembrane prostate androgen-induced protein gene suppressed triple-negative breast cancer cell proliferation. Med J Indones [Internet]. 2017Nov.27 [cited 2019Aug.19];26(3):178-82. Available from: http://mji.ui.ac.id/journal/index.php/mji/article/view/1823
Section
Basic Medical Research