TY - JOUR
T1 - Role of TGF-β1 in human breast cancer stem cells
AU - Hariyanto, Narendra Ichiputra
AU - Purwandhita, Rini Putri
AU - Syahrani, Resda Akhra
AU - Louisa, Melva
AU - Wanandi, Septelia Inawati
N1 - Funding Information:
Funding Disclosure: This research was funded by Master Thesis Grant (Hibah Tesis Magister) 2019 from Ministry of Research, Technology, and Higher Education of the Republic of Indonesia.
Publisher Copyright:
© 2021 Pakistan Medical Association. All rights reserved.
PY - 2021/2/1
Y1 - 2021/2/1
N2 - OBJECTIVE: To investigate the auto-induction of transforming growth factor-b1 (TGF-β1) in breast cancer stem cells (BCSCs) and its effect on cell viability and stemness. Methods: Human BCSCs (aldehyde dehydrogenase positive; ALDH+) were grown in serum-free Dulbecco's Modified Eagle Medium/Nutrient Mixture F12 (DMEM/F12) and treated for periods of 1, 2 and 4 hours with 0.1 ng/ml recombinant human TGF-β1 protein (rhTGF-β1). The medium was then replaced with serum-free DMEM/F12 without rhTGF-β1 for 24 hours. Cell viability was determined using a trypan blue exclusion assay. Type 1 TGF-β receptor (TβR1), TGF-β1, octamer-binding transcription factor 4 (OCT4) and aldehyde dehydrogenase 1 family member A1 (ALDH1A1) messenger RNA (mRNA) expression levels were analysed using quantitative real-time reverse-transcriptase polymerase chain reaction (RT-qPCR). The TGF-β protein level in the culture medium was determined using an enzyme-linked immunosorbent assay (ELISA). RESULTS: The expression levels of rhTGF-β1, TGF-β1 and TβR1 mRNA significantly increased in BCSCs compared to control after treatment for 1 and 2 hours but decreased after 4 hours. This is in line with alteration of stemness gene, OCT4 and ALDH1A1 mRNA expressions. However, the secretion of newly synthesised TGF-β1 significantly increased after 2 hours. In contrast, viable BCSCs decreased after 1 hour and then gradually increased 2.7 times compared to control after 4 hours. CONCLUSIONS: TGF-β1 treatment in low concentration and for short period of time triggers its auto-induction in BCSCs, leading to increased cell viability and stemness gene expression via autocrine signalling.
AB - OBJECTIVE: To investigate the auto-induction of transforming growth factor-b1 (TGF-β1) in breast cancer stem cells (BCSCs) and its effect on cell viability and stemness. Methods: Human BCSCs (aldehyde dehydrogenase positive; ALDH+) were grown in serum-free Dulbecco's Modified Eagle Medium/Nutrient Mixture F12 (DMEM/F12) and treated for periods of 1, 2 and 4 hours with 0.1 ng/ml recombinant human TGF-β1 protein (rhTGF-β1). The medium was then replaced with serum-free DMEM/F12 without rhTGF-β1 for 24 hours. Cell viability was determined using a trypan blue exclusion assay. Type 1 TGF-β receptor (TβR1), TGF-β1, octamer-binding transcription factor 4 (OCT4) and aldehyde dehydrogenase 1 family member A1 (ALDH1A1) messenger RNA (mRNA) expression levels were analysed using quantitative real-time reverse-transcriptase polymerase chain reaction (RT-qPCR). The TGF-β protein level in the culture medium was determined using an enzyme-linked immunosorbent assay (ELISA). RESULTS: The expression levels of rhTGF-β1, TGF-β1 and TβR1 mRNA significantly increased in BCSCs compared to control after treatment for 1 and 2 hours but decreased after 4 hours. This is in line with alteration of stemness gene, OCT4 and ALDH1A1 mRNA expressions. However, the secretion of newly synthesised TGF-β1 significantly increased after 2 hours. In contrast, viable BCSCs decreased after 1 hour and then gradually increased 2.7 times compared to control after 4 hours. CONCLUSIONS: TGF-β1 treatment in low concentration and for short period of time triggers its auto-induction in BCSCs, leading to increased cell viability and stemness gene expression via autocrine signalling.
KW - Transforming Growth Factor beta1, Breast Neoplasms, Neoplastic Stem Cells, Octamer Transcription Factor-3, Aldehyde Dehydrogenase.
UR - http://www.scopus.com/inward/record.url?scp=85103745958&partnerID=8YFLogxK
M3 - Article
C2 - 33785948
AN - SCOPUS:85103745958
SN - 0030-9982
VL - 71 2)
SP - S84-S89
JO - JPMA. The Journal of the Pakistan Medical Association
JF - JPMA. The Journal of the Pakistan Medical Association
IS - 2
ER -
