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

Authors

  • 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

DOI:

https://doi.org/10.13181/mji.v26i3.1823

Keywords:

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

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.

Downloads

Download data is not yet available.

References

Ng CH, Pathy NB, Taib NA, Teh YC, Mun KS, Amiruddin A, et al. Comparison of breast cancer in Indonesia and Malaysia-A Clinico-Pathological study between Dharmais Cancer Center Jakarta and University Malaya Medical Center, Kuala Lumpur. Asian Pacific J Cancer Prev. 2011;12:2943-6.

Singha PK, Pandeswara S, Geng H, Lan R, Venkatachalam MA, Saikumar P. TGF-β induced TMEPAI/PMEPA1 inhibits canonical Smad signaling through R-Smad Sequestration and promotes non-canonical PI3K/Akt signaling by reducing PTEN in triple negative breast cancer. Genes&Cancer. 2014;5(9-10):320-36. https://doi.org/10.18632/genesandcancer.30

Foulkes WD, Smith IE, Reis-Fielho JS. Triple negative breast cancer. N Eng J Med. 2010;363:1938-48. https://doi.org/10.1056/NEJMra1001389

Oâ??Reilly EA, Gubbins L, Sharma S, Tully R, Guang MHZ, Weiner-Gorzel K, et al. The fate of chemoresistance in triple negative breast cancer (TNBC). BBA Clinical 3. 2015;3:257-75. https://doi.org/10.1016/j.bbacli.2015.03.003

Gyorffy B, Lanczky A, Eklund AC, Denkert C, Budczies J, Li Q, Szallasi Z. An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1809 patients. Breast Cancer Res Treat. 2010 Oct;123(3):725-31. doi: 10.1007/s10549-009-0674-9

Singha PK, Yeh IT, Venkatachalam MA, Saikumar P. TGF-v-Inducible gene TMEPAI converts TGF- v from a tumor suppressor to a tumor promoter in breast cancer. Cancer Res. 2009;70:6377–83. https://doi.org/10.1158/0008-5472.CAN-10-1180

Watanabe Y, Itoh S, Goto T, Ohnishi E, Inamitsu M, Itoh F, et al. TMEPAI, a transmembrane TGF-v-inducible protein, sequesters Smad proteins from active participation in TGF-v signaling. Mol Cell. 2010;37:123–34. https://doi.org/10.1016/j.molcel.2009.10.028

Ngunyen TTV, Watanabe Y, Shiba A, Noguchi M, Itoh S, Kato M. TMEPAI/PMEPA1 enhances tumorigenic activities in lung cancer cells. Cancer Sci. 2014;105(3):334–41. https://doi.org/10.1111/cas.12355

Wardhani BWK, Puteri MU, Watanabe Y, Louisa M, Setiabudy R, et al. TMEPAI genome editing in triple negative breast cancer cells. Med J Indones. 2016;26:14-8. https://doi.org/10.13181/mji.v26i1.1871

Roth V. 2006 Doubling Time Computing [Internet]. [cited 2016]. Available from: http://www.doubling-time.com/compute.php

Azami S, Vo Nguyen T, Watanabe Y, Kato M. Cooperative induction of transmembrane prostate androgeninduced protein TMEPAI/PMEPA1 by transforming growth factor-v and epidermal growth factor signaling. Biochem Biophy Res Commun. 2015;456:580–5. https://doi.org/10.1016/j.bbrc.2014.11.107

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 2024Dec.1];26(3):178-82. Available from: https://mji.ui.ac.id/journal/index.php/mji/article/view/1823

Issue

Section

Basic Medical Research
Abstract viewed = 1719 times

Most read articles by the same author(s)

1 2 3 > >>