HDAC2 and PCNA expression is correlated to decreasing of endoxifen sensitivity in human breast cancer stem cells ALDH+

  • Syarifah Dewi
  • Mohamad Sadikin
  • Muchlis Ramli
  • Septelia Inawati Wanandi
Keywords: human breast cancer stem cells, viability, HDAC2, PCNA, endoxifen

Abstract

Latar belakang: Sel punca kanker payudara (breast cancer stem cells/BCSC) adalah subpopulasi sel kanker yang memiliki kemampuan menghasilkan tumor baru dan bersifat seperti sel punca. Penelitian kami sebelumnya menggunakan jaringan kanker payudara mengungkapkan bahwa ekspresi gen histone deacetylase 2 (HDAC2) dan proliferating cell nuclear antigen (PCNA) ditemukan perbedaan signifikan setelah terapi neoajuvan hormon dan kemoterapi. Penelitian ini bertujuan untuk menganalisis hubungan antara ekspresi HDAC2 dan PCNA dengan kelangsungan hidup sel punca kanker payudara dengan penanda aldehyde dehydrogenase + (ALDH+) yang diberi perlakuan endoksifen.


Metode: Sampel adalah BCSC primer manusia ALDH+ yang diberi perlakuan endoksifen 4 uM masingmasing selama 2, 4, 6, 8, 10, 12, 14 hari. Viabilitas sel dilihat dengan menggunakan trypan blue dan ekspresi mRNA HDAC2 dan PCNA ditentukan menggunakan qRT-PCR.


Hasil: Viabilitas BCSCs ALDH + menurun setelah 2 sampai 4 hari pemberian endoksifen. Pada periode ini juga didapatkan ekspresi mRNA HDAC2 dan PCNA mengalami penurunan. Tetapi setelah pemberian endoksifen selama 8 hari, viabilitas BCSCs ALDH + mengalami peningkatan dan ditemukan peningkatan yang signifikan pada hari ke-14 pemberian endoksifen. Ekspresi mRNA HDAC2 dan PCNA juga menunjukkan peningkatan mulai pada hari ke-8 dan terus meningkat hingga hari ke-14 pemberian endoksifen. Penelitian ini menunjukkan pola yang sama antara ekspresi mRNA HDAC2 dan PCNA dan viabilitas sel.


Kesimpulan: Induksi endoksifen yang lama menurunkan sensitivitas efek endoksifen pada BCSC manusia dan ekspresi HDAC2 dan PCNA berkorelasi dengan viabilitas BCSC manusia setelah induksi endoksifen. (Health Science Journal of Indonesia 2019;10(2):77-81)


Kata kunci: sel punca kanker payudara, viabilitas sel, HDAC2, PCNA, endoksifen

 

Abstract


Background: Breast cancer stem cells (BCSCs) are subpopulation of cancer cells that has the ability to generate new tumor and similar properties to stem cell. Our previous study using breast cancer patients revealed that gene expression of histone deacetylase 2 (HDAC2) and proliferating cell nuclear antigen (PCNA) were significantly altered after neoadjuvant hormone and chemotherapy. This study aimed to analyze the correlation between HDAC2 and PCNA expressions with the viability of breast cancer stem cells aldehyde dehydrogenase + (BCSC ALDH+) treated by endoxifen.


Method: Samples are human primary BCSCs ALDH+ that treated with 4 uM of endoxifen for 2, 4, 6, 8, 10, 12, 14 days, respectively. Cell viability was measured using trypan blue exclusion assay and the mRNA expressions of HDAC2 and PCNA were determined using qRT-PCR.


Results: The viability of BCSCs ALDH+ was decreased after 2 days until 4 days-endoxifen treatment. It also demonstrated that mRNA expression of HDAC2 and PCNA were decreased in this period. But after 8 days endoxifen treatment, the viability of BCSCs ALDH+ was increased. The increasing of viability was higher in 14 days-endoxifen treatment. The mRNA expression of HDAC2 and PCNA also showed increasing begin on 8 days and continued to increase until 14-days endoxifen treatment. We found a similar pattern between HDAC2 and PCNA expression and cell viability.


Conclusion: Prolonge endoxifen treatment decrease sensitivity of endoxifen effect in human BCSC and the expression of HDAC2 and PCNA are correlated to human BCSCs viability after endoxifen treatment. (Health Science Journal of Indonesia 2019;10(2):77-81)


Keywords: human breast cancer stem cells, viability, HDAC2, PCNA, endoxifen

References

Kim HJ, Bae SC. Histone deacetylase inhibitors: molecular mechanisms of action and clinical trials as anti cancer drugs. Am J Transl Res. 2011;3(2):166–79.

Edward S, Minoru Y. Erasers of histone acetylation: the histone deacetylase enzymes. Cold Spring Harbor Perspectives in Biology. Cold Spring Harb Perspect Med. 2014;6(4):a018713.

Qiu X, Mei J, Yin J, Wang H, Wang J, Xie M. Correlation analysis between expression of PCNA, Ki- 67 and COX-2 and X-ray features in mammography in breast cancer. Oncol Lett. 2017;14(3):2912–8.

Wang X, Wang D, Yuan N, Liu F, Wang F, Wang B, et al. The prognostic value of PCNA expression in patients with osteosarcoma: A meta-analysis of 16 studies. Medicine. 2017;96(41):e8254–e8254.

Wu X, Hawse JR, Subramaniam M, Goetz MP, Ingle JN, Spelsberg TC. The tamoxifen metabolite, endoxifen, is a potent antiestrogen that targets estrogen receptor a for degradation in breast cancer cells. Cancer Res. 2009;69(5):1722–7.

Gangopadhyay S, Nandy A, Hor P, Mukhopadhyay A. Breast cancer stem cells: a novel therapeutic target. Clin Breast Cancer. 2013;13(1):7–15.

Wanandi SI, Dewi S, Pramana S, Karsono R. The alteration of p53-pathway gene expression in advanced breast cancer after neoadjuvant chemo- and hormone therapy. Ann Oncol. 2016;27(9):ix38.

Dewi S, Karsono R, Pramana S, Wanandi SI. Expression profile of stem cell pathway genes in patients with advanced breast cancer after neoadjuvant therapy. J Phys Conf Ser. 2018;1073(3).

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2( Delta Delta C(T)) Method. Methods. 2001;25(4):402–8.

Witt AE, Lee C, Lee TI, Azzam DJ, Wang B, Caslini C, et al. Identification of a cancer stem cell-specific function for the histone deacetylases, HDAC1 and HDAC7, in breast and ovarian cancer. Nat Publ Gr. 2016;36(12):1707–20.

Li Y, Seto E. HDACs and HDAC inhibitors in cancer development and therapy. Cold Spring Harb Perspect Med. 2016 Oct 3;6(10):a026831.

Ropero S, Esteller M. The role of histone deacetylases (HDACs) in human cancer. Mol Oncol. 2007;1(1):19– 25.

Mann M, Cortez V, Vadlamudi RK. Epigenetics of estrogen receptor signaling: Role in hormonal cancer progression and therapy. Cancers (Basel). 2011;3:1691–707.

Begicevic R-R, Falasca M. ABC transporters in cancer stem cells: beyond chemoresistance. Int J Mol Sci. 2017;18(11):2362.

Bhaskara S, Jacques V, Rusche JR, Olson EN, Cairns BR, Chandrasekharan MB. Histone deacetylases 1 and 2 maintain S-phase chromatin and DNA replication fork progression. Epigenetics Chromatin. 2013;6(1):27.

Bhaskara S. Histone deacetylases 1 and 2 regulate DNA replication and DNA repair: potential targets for genome stability-mechanism-based therapeutics for a subset of cancers. Cell Cycle. 2015;14(12):1779–85.

Zhao H, Yu Z, Zhao L, He M, Ren J, Wu H, et al. HDAC2 overexpression is a poor prognostic factor of breast cancer patients with increased multidrug resistance-associated protein expression who received anthracyclines therapy. Jpn J Clin Oncol. 2016;46(10):893–902.

Morell-Quadreny L, Clar-Blanch F, Fenollosa-Enterna B, Perez-Bacete M, Martinez-Lorente A, Llombart-Bosch A. Proliferating cell nuclear antigen (PCNA) as a prognostic factor in renal cell carcinoma. Anticancer Res. 1998;18(1B):677–82.

Moldovan GL, Pfander B, Jentsch S. PCNA, the maestro of the replication fork. Cell. 2007;129(4):665–79.

Published
2019-12-31
How to Cite
Dewi, S., Sadikin, M., Ramli, M., & Wanandi, S. (2019). HDAC2 and PCNA expression is correlated to decreasing of endoxifen sensitivity in human breast cancer stem cells ALDH+. Health Science Journal of Indonesia, 10(2), 77-81. https://doi.org/10.22435/hsji.v12i2.2449
Section
Articles