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

Bantari W.K. Wardhani, Meidi U. Puteri, Yukihide Watanabe, Melva Louisa, Rianto Setiabudy, Mitsuyasu Kato



DOI: http://dx.doi.org/10.13181/mji.v26i3.1823

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.


Keywords


proliferation; TGF-β; TMEPAI; triple negative breast cancer

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References


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Copyright (c) 2017 Bantari W.K. Wardhani, Meidi Utami Puteri, Yukihide Watanabe, Melva Louisa, Rianto Setiabudy, Mitsuyasu Kato

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