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Journal : The Indonesian Biomedical Journal

Dedifferentiation of MCF-7 Breast Cancer Continuous Cell Line, Development of Breast Cancer Stem Cells (BCSCs) Enriched Culture and Biomarker Analysis Ami Ashariati Prayogo; Andi Yasmin Wijaya; Winona May Hendrata; Steven Sheng Looi; Reny I’tishom; Lukman Hakim; Fedik Abdul Rantam; I Ketut Sudiana; Abdurachman Abdurachman
The Indonesian Biomedical Journal Vol 12, No 2 (2020)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v12i2.977

Abstract

BACKGROUND: Cancer stem cells (CSCs) eradication might serve as a robust approach for cancer eradication. MCF-7 as breast cancer continuous cell line is known to contain breast CSCs (BCSCs) for its capability to maintain its original tumor population. CSCs enriched culture is a fundamental tool for CSCs targeted therapy development. Effective and unsophisticated CSCs dedifferentiation protocol for producing CSCs enriched culture is needed.METHODS: MCF-7 cells were cultured initially in Dulbecco's Modified Eagle Medium (DMEM) low glucose medium then changed to DMEM:F12. Serum starvation was performed during each medium refreshment gradually with fetal bovine serum (FBS) concentration of 10%, 5%, 2.5% until reaching 1% FBS concentration. Stable MCF-7 culture was then adapted to serum free culture system, containing DMEM:F12, epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and B27 supplement as dedifferentiation protocol for 18 days. Cluster of differentiation (CD)44 and CD24 double staining immunocytochemistry was performed to evaluate cell stemness.RESULTS: The population of cells expressing BCSCs markers (CD44+/CD24low) in non-adherent single cells subpopulation was significantly increased after the dedifferentiation procedure (70.39%) compared to control groups (0.71%) (p<0.05). In contrast, the expression of BCSCs marker in adherent single cells subpopulation and for both adherent and non-adherent mammosphere the BCSCs markers showed a stable expression.CONCLUSION: BCSCs enrichment of breast cancer cell cultures from MCF-7 breast cancer cell line can be performed. Breast cancer cell plasticity is observed during the dedifferentiation protocol. Development of dedifferentiation inducing protocols can serve as an important foundation for breast cancer therapy development through BCSCs elimination.KEYWORDS: breast neoplasms, cell line, dedifferentiation, immunohistochemistry, neoplastic stem cells
Inhibition of Neurogenesis and Induction of Glial Scar Formation by Neuroinflammation Following Ischemic Stroke: Evaluation of BDNF, GFAP, HMGB1 and TNF-α Expressions Aris Widayati; Fedik Abdul Rantam; Abdulloh Machin; Wibi Riawan
The Indonesian Biomedical Journal Vol 17, No 1 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i1.3439

Abstract

BACKGROUND: Ischemic stroke remains as a major health problem and one important process in Ischemic Stroke is neuroinflammation which has a principal role to maintain the balance of neurogenesis and neurodegeneration process in the brain. Neuroinflammation can lead to glial scar and inhibit neurogenesis processes which is needed for recovery neuron function. This study was conducted to observe the role of high mobility group box 1 (HMGB1) and tumor necrosis factor-α (TNF-α) as neuroinflammation markers to glial fibrillary acidic protein (GFAP) as glial scar marker and to brain-derived neurotrophic factor (BDNF) as neurogenesis marker in brain tissue following ischemic stroke.METHODS: Fifteen male Wistar rats were randomized to three groups; sham group, rats receiving occlusion and terminated 180 minutes later (group A), and rats receiving occlusion and terminated after 7 days (group B). Expressions of BDNF and BDNF mRNA were examined using immunohistochemistry (IHC) and Reverse Transcription Polymerase Chain Reaction (RT-PCR), respectively. While GFAP, HMGB1, TNF-α were assessed using IHC.RESULTS: Expression of BDNF was found lower in group A and group B than in sham group (5.20±1.924, 5.00±1.581, and 7.80±1.304, respectively; p=0.032). Expression of BNDF mRNA was found lower in group A and B than in sham group as well. While expression of GFAP was found higher in group A and B than in sham group (9.60±1.517, 11.40±2.074, and 5.20±1.48, respectively; p=0.000). Higher level of HMGB1 and TNF-α expressions were also found to in group A and group B than in sham group (9.3±1.528, 11.67±1.528, and 2.00±1.000, respectively; p=0.000 for HMGB1 and 6.33±1.155, 9.33±1.528, and 3.00±1.000, respectively; p=0.002 for TNF-α).CONCLUSION: The presence of low BDNF levels and high levels of GFAP, HMGB1 and TNF-α markers, possibly reflects inhibition of the neurogenesis process by neuroinflammation, and induced glial scar formation in ischemic stroke conditions after than 180 hours until 7 days. KEYWORDS: ischemic stroke, BDNF, GFAP, TNF-α, HMGB1
Co-Authors . Helen . Mulyani . Purwati . PURWATI . Suwarno A'la, Rofiqul Abdurachman Abdurachman Ahmad Chusnu Romdhoni Aksono HP., Eduardus Bimo Amaq Fadholly Ami Ashariati Amrullah, Amirul Muslim Andi Yasmin Wijaya Andriyanto Andriyanto Anindya, Afifatus Annise Proboningrat Ardhanella, Sellianova Arif Nur Muhammad Ansori Aris Widayati Aswin, Ahmad Aucky Hinting Aulia Andi Mustika Azahro, Aisyah Bambang Sektiari Lukiswanto Boedi Setiawan Candra Bumi Cita Rosita Sigit Prakoeswa Damayanti Tinduh Desi Sandra Sari Diah Nugrahani Pristihadi Didik Handijatno Diyantoro Diyantoro, Diyantoro Dwikora Novembri Utomo Emmanuel Djoko Poetranto ENDANG RETNOWATI KUSUMOWIDAGDO Endang Suprihati ERMA SAFITRI Ernawati, Rahaju Ernawati, Rahayu Ernie Maduratna Estoepangestie, Agnes Theresia Soelih Ferdiansyah Ferdiansyah Mahyudin Gilang Maulana Putra HAK HOTTA Hani Plumeriastuti Hari Basuki Notobroto Harijani, Nenny Helen Susilowati, Helen Henry Dominica Heri Suroto HERNOMO ONTOSENO KUSUMOBROTO I'tishom, Reny Ika WAHYUNI IMAM MUSTOFA Imam Mustofa Indah Rahmawati Ira Sari Yudaniayanti Iwan Sahrial Hamid Juniastuti Juniastuti Kadek Rachmawati Khairullah, Aswin Rafif Kharisma, Viol Dhea Kirana, Maha Kuncorojakti, Suryo Kusnoto Kusnoto, Kusnoto Laba Mahaputra Lina Noviyanti Sutardi Lucia Tri Suwanti, Lucia Tri Lukman Hakim Machin, Abdulloh MARIA INGE LUSIDA Mas’ud Hariadi Mouli Edward Mufasirin Mustofa Helmi Effendi Mutiarasari, Nonie Olivia Adia Nasronudin Nasronudin Ni Ketut Suwiti Nidom, Chairul Anwar Nunuk Dyah Retno Lastuti Nusdianto Triakoso Purwati Purwati Putih, Nurvita R. Heru Prasetyo Rahaju Ernawati Raharjo, Dadik Rahmahani, Jola Ratih Ratnasari Ratna Damayanti Retno Pudji Rahayu Ria Hanistya Rina Mutya Suzliana Romaniyanto Romaniyanto Romaniyanto Romy Muhammad Dary Mufa S.Pd. M Kes I Ketut Sudiana . Santoso, Kuncoro Puguh SOETJIPTO . Sri Hernawati Sri Pantja Madyawati Sri Wigati Mardi Steven Sheng Looi Suwarno . Suwarno Suwarno Suwarno Suwarno Suzanita Utama Tacharina, Martia Rani TAKAKO UTSUMI Tristiana Erawati Tyasningsih, Wiwiek Venri Novryantoro Wibi Riawan Widiyatno, Thomas Valentinus Widji Soeratri Widjiati . Widjiati Widjiati Widodo Ario Kentjono Wijaya, Andi Yasmin Winona May Hendrata Wulandari, Innah Yoes Priyatna Dachlan YOSHIHIKO YANO YOSHITAKE HAYASHI Yulanda Antonius Yusuf, Muhtarum Zahro, Vida Seanita Zayyin Dinana