Knock-out transmembrane prostate androgen-induced protein gene suppressed triple-negative breast cancer cell proliferation
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.
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
O’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-β-Inducible gene TMEPAI converts TGF- β 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-β-inducible protein, sequesters Smad proteins from active participation in TGF-β 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 androgen-induced protein TMEPAI/PMEPA1 by transforming growth factor-β and epidermal growth factor signaling. Biochem Biophy Res Commun. 2015;456:580–5. https://doi.org/10.1016/j.bbrc.2014.11.107
Copyright (c) 2017 Bantari W.K. Wardhani, Meidi U. Puteri, Yukihide Watanabe, Melva Louisa, Rianto Setiabudy, Mitsuyasu Kato
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with Medical Journal of Indonesia agree to the following terms:
- Authors retain copyright and grant Medical Journal of Indonesia right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial License that allows others to remix, adapt, build upon the work non-commercially with an acknowledgment of the work’s authorship and initial publication in Medical Journal of Indonesia.
- Authors are permitted to copy and redistribute the journal's published version of the work non-commercially (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in Medical Journal of Indonesia.