Öz
Amaç: Çalışmada, dünya genelinde en yaygın kanser tiplerinden biri olan meme kanserinde, apoptoz ile ilişkili genlerin anastaz sürecindeki ifadesel değişimlerinin ve potansiyel rollerinin tanımlanması amaçlanmıştır.
Gereç ve Yöntem: Farklı tipteki meme kanseri hücre hatları (MCF7 ve MDA-MB-231), meme kanseri kök hücreleri ve sağlıklı meme hücre hattı (MCF10A) kullanıldı. Apoptotik ve anastatik hücre yüzdeleri, akış sitometrisi aracılığıyla Annexin V testi ile belirlendi. Apoptotik hücrelere göre anastatik hücrelerin gen ekspresyon değişimleri qRT-PCR ve 2-ΔΔCt yöntemi ile belirlendi. Anlamlı değişim gösteren genlerin görev aldıkları yolak ve biyolojik süreçler STRING v11.5 veri tabanı kullanılarak belirlendi.
Bulgular: Tüm hücre gruplarında etanol uygulaması sonucu kontrole göre apoptoz yüzdesinin arttığı ve apoptozu uyaran etmenin uzaklaştırılmasıyla apoptotik hücre yüzdesinin azaldığı belirlendi. Kontrol, apoptoz ve anastaz grupları arasında apoptotik hücre yüzdesindeki değişim en fazla MCF7 hücrelerinde belirlendi. Uyumlu şekilde bu hücre hattında en fazla sayıda gende ifade değişimi belirlendi. CASP7 ve APAF1 genleri tüm hücre hatlarında ekspresyon azalışı sergiledi. Tüm hücre gruplarında anastazın sistein tipi endopeptidaz aktivitesini (GO_ID: 0043027) ve ilaç direnci ilişkili yolakları (KEGG_ID: hsa01524) ortak şekilde etkilediği belirlendi.
Sonuç: Hücrelerde anastaz fenomeninin apoptoz düzenleyici mekanizmalar ile etkileşiminin tanımlanması, hem sağlıklı hücrelerin onkojenik dönüşümünün hem de kanserde ilaç direnci mekanizmalarının aydınlatılabilmesi açısından önem taşımaktadır.
Anahtar Kelimeler
Kaynakça
- Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71(3):209-249. doi:10.3322/caac.21660
- Kusoglu A, Goker Bagca B, Ozates Ay NP, Gunduz C, Biray Avci C. Telomerase inhibition regulates EMT mechanism in breast cancer stem cells. Gene. Published online 2020. doi:10.1016/j.gene.2020.145001
- Galluzzi L, Vitale I, Aaronson SA, et al. Molecular mechanisms of cell death: Recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death Differ. Published online 2018. doi:10.1038/s41418-017-0012-4
- Tang HL, Tang HM, Mak KH, et al. Cell survival, DNA damage, and oncogenic transformation after a transient and reversible apoptotic response. Mol Biol Cell. Published online 2012. doi:10.1091/mbc.E11-11-0926
- INCEBOZ M, GOKER BAGCA B, CANER A, GÜNDÜZ C. Anastasis in Glioblastoma, Brain Cancer Stem, and Brain Stem Cells. J Basic Clin Heal Sci. Published online 2021. doi:10.30621/jbachs.854986
- Tang HM, Tang HL. Anastasis: Recovery from the brink of cell death. R Soc Open Sci. Published online 2018. doi:10.1098/rsos.180442
- Szklarczyk D, Gable AL, Nastou KC, et al. The STRING database in 2021: Customizable protein-protein networks, and functional characterization of user-uploaded gene/measurement sets. Nucleic Acids Res. Published online 2021. doi:10.1093/nar/gkaa1074
- Campbell KJ, Tait SWG. Targeting BCL-2 regulated apoptosis in cancer. Open Biol. Published online 2018. doi:10.1098/rsob.180002
- Liu Y, Zuo H, Wang Y, et al. Ethanol promotes apoptosis in rat ovarian granulosa cells via the Bcl-2 family dependent intrinsic apoptotic pathway. Cell Mol Biol. Published online 2018. doi:10.14715/cmb/2018.64.1.21
- Soengas MS, Alarcón RM, Yoshida H, et al. Apaf-1 and caspase-9 in p53-dependent apoptosis and tumor inhibition. Science (80- ). Published online 1999. doi:10.1126/science.284.5411.156
- Bakhshoudeh M, Mehdizadeh K, Hosseinkhani S, Ataei F. Upregulation of apoptotic protease activating factor-1 expression correlates with anti-tumor effect of taxane drug. Med Oncol. Published online 2021. doi:10.1007/s12032-021-01532-8
- Siegmund D, Mauri D, Peters N, et al. Fas-associated Death Domain Protein (FADD) and Caspase-8 Mediate Up-regulation of c-Fos by Fas Ligand and Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) via a FLICE Inhibitory Protein (FLIP)-regulated Pathway. J Biol Chem. Published online 2001. doi:10.1074/jbc.M100444200
- Hsu H, Xiong J, Goeddel Dv. The TNF receptor 1 associated protein TRADD signals cell death and NF kappa B activation. Cell. Published online 1995.
- Pobezinskaya YL, Liu Z. The role of TRADD in death receptor signaling. Cell Cycle. Published online 2012. doi:10.4161/cc.11.5.19300
- Chandler JM, Cohen GM, MacFarlane M. Different subcellular distribution of caspase-3 and caspase-7 following Fas-induced apoptosis in mouse liver. J Biol Chem. Published online 1998. doi:10.1074/jbc.273.18.10815
- Scott FL, Denault JB, Riedl SJ, Shin H, Renatus M, Salvesen GS. XIAP inhibits caspase-3 and -7 using two binding sites: Evolutionary conserved mechanism of IAPs. EMBO J. Published online 2005. doi:10.1038/sj.emboj.7600544
- Polyak K, Xia Y, Zweier JL, Kinzler KW, Vogelstein B. A model for p53-induced apoptosis. Nature. Published online 1997. doi:10.1038/38525
Öz
Aim: This study aimed to define the expression changes and potential roles of the apoptosis-related genes in the anastasis process in breast cancer, which is one of the most common cancer types.
Materials and Methods: Different types of breast cancer cell lines (MCF7 and MDA-MB-231), breast cancer stem cells, and healthy breast cell line (MCF10A) were used. Apoptotic and anastatic cell percentages were determined by the Annexin V test and flow cytometry. Gene expression changes in anastatic cells compared to apoptotic cells were determined by the qRT-PCR and 2-ΔΔCt method. The pathways and biological processes of genes that show significant changes were determined using the STRING v11.5 database.
Results: In all cell groups, it was determined that the percentage of apoptosis increased as a result of ethanol application, and the percentage of apoptotic cells decreased with the removal of the apoptosis-inducing factor. The change in the percentage of apoptotic cells between the control, apoptosis, and anastasis groups was determined the most in MCF7 cells. Consistently, expression changes were determined in the largest number of genes in this cell line. CASP7 and APAF1 genes downregulated in all cell lines. In all cell groups, it was determined that anastasis affects Cysteine-type endopeptidase activity involved in execution phase of apoptosis (GO_ID: 0043027), and drug resistance-related pathways (KEGG_ID: hsa01524).
Conclusion: The definition of the interaction of the anastasis phenomenon in cells with apoptosis regulatory mechanisms is important in terms of elucidating both the oncogenic transformation of healthy cells and the mechanisms of drug resistance in cancer.
Anahtar Kelimeler
Kaynakça
- Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71(3):209-249. doi:10.3322/caac.21660
- Kusoglu A, Goker Bagca B, Ozates Ay NP, Gunduz C, Biray Avci C. Telomerase inhibition regulates EMT mechanism in breast cancer stem cells. Gene. Published online 2020. doi:10.1016/j.gene.2020.145001
- Galluzzi L, Vitale I, Aaronson SA, et al. Molecular mechanisms of cell death: Recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death Differ. Published online 2018. doi:10.1038/s41418-017-0012-4
- Tang HL, Tang HM, Mak KH, et al. Cell survival, DNA damage, and oncogenic transformation after a transient and reversible apoptotic response. Mol Biol Cell. Published online 2012. doi:10.1091/mbc.E11-11-0926
- INCEBOZ M, GOKER BAGCA B, CANER A, GÜNDÜZ C. Anastasis in Glioblastoma, Brain Cancer Stem, and Brain Stem Cells. J Basic Clin Heal Sci. Published online 2021. doi:10.30621/jbachs.854986
- Tang HM, Tang HL. Anastasis: Recovery from the brink of cell death. R Soc Open Sci. Published online 2018. doi:10.1098/rsos.180442
- Szklarczyk D, Gable AL, Nastou KC, et al. The STRING database in 2021: Customizable protein-protein networks, and functional characterization of user-uploaded gene/measurement sets. Nucleic Acids Res. Published online 2021. doi:10.1093/nar/gkaa1074
- Campbell KJ, Tait SWG. Targeting BCL-2 regulated apoptosis in cancer. Open Biol. Published online 2018. doi:10.1098/rsob.180002
- Liu Y, Zuo H, Wang Y, et al. Ethanol promotes apoptosis in rat ovarian granulosa cells via the Bcl-2 family dependent intrinsic apoptotic pathway. Cell Mol Biol. Published online 2018. doi:10.14715/cmb/2018.64.1.21
- Soengas MS, Alarcón RM, Yoshida H, et al. Apaf-1 and caspase-9 in p53-dependent apoptosis and tumor inhibition. Science (80- ). Published online 1999. doi:10.1126/science.284.5411.156
- Bakhshoudeh M, Mehdizadeh K, Hosseinkhani S, Ataei F. Upregulation of apoptotic protease activating factor-1 expression correlates with anti-tumor effect of taxane drug. Med Oncol. Published online 2021. doi:10.1007/s12032-021-01532-8
- Siegmund D, Mauri D, Peters N, et al. Fas-associated Death Domain Protein (FADD) and Caspase-8 Mediate Up-regulation of c-Fos by Fas Ligand and Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) via a FLICE Inhibitory Protein (FLIP)-regulated Pathway. J Biol Chem. Published online 2001. doi:10.1074/jbc.M100444200
- Hsu H, Xiong J, Goeddel Dv. The TNF receptor 1 associated protein TRADD signals cell death and NF kappa B activation. Cell. Published online 1995.
- Pobezinskaya YL, Liu Z. The role of TRADD in death receptor signaling. Cell Cycle. Published online 2012. doi:10.4161/cc.11.5.19300
- Chandler JM, Cohen GM, MacFarlane M. Different subcellular distribution of caspase-3 and caspase-7 following Fas-induced apoptosis in mouse liver. J Biol Chem. Published online 1998. doi:10.1074/jbc.273.18.10815
- Scott FL, Denault JB, Riedl SJ, Shin H, Renatus M, Salvesen GS. XIAP inhibits caspase-3 and -7 using two binding sites: Evolutionary conserved mechanism of IAPs. EMBO J. Published online 2005. doi:10.1038/sj.emboj.7600544
- Polyak K, Xia Y, Zweier JL, Kinzler KW, Vogelstein B. A model for p53-induced apoptosis. Nature. Published online 1997. doi:10.1038/38525
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Sağlık Kurumları Yönetimi |
| Bölüm | Araştırma Makaleleri |
| Yazarlar | |
| Yayımlanma Tarihi | 12 Eylül 2022 |
| Gönderilme Tarihi | 22 Şubat 2022 |
| Yayımlandığı Sayı | Yıl 2022Cilt: 61 Sayı: 3 |
