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Pengaruh Pemberian TGF-Β1 Rekombinan Manusia pada Sel Punca Kanker Payudara ALDH+ terhadap Ekspresi Marker Kepuncaan Melalui Pensinyalan Autokrin TGF-Β1 = The Impact of Human TGF-Β1 Recombinant Protein Treatment to BCSC ALDH+ on The Expression of Stemness Marker Through TGF-Β1 Autocrine Signaling

Narendra Ichiputra Hariyanto; Septelia Inawati Wanandi, supervisor; Melva Louisa, supervisor; Siregar, Nurjati Chairani, examiner; Vivian Soetikno, examiner (Fakultas Kedokteran Universitas Indonesia, 2020)

 Abstrak

Latar Belakang: Breast Cancer Stem Cells (BCSC) merupakan populasi sel
kanker payudara yang mempunyai sifat sel punca. BCSC menjaga stabilitas tumor
dengan menginisiasi pembentukan populasi sel kanker baru serta memberikan
kekebalan terhadap terapi. BCSC dapat berinteraksi dengan lingkungan mikro
tumor yang melepaskan berbagai sitokin dan growth factor, termasuk TGF-β1.
Melalui mekanisme autoinduksi, TGF-β1 dapat meningkatkan produksi TGF-β1
endogen dan pensinyalan autokrinnya. Pensinyalan autokrin TGF-β1 dapat
meningkatkan pengaruh tumor promotor TGF-β1 dalam perkembangan kanker
melalui penguatan karakter kepuncaan dan sifat tumorigenik. Penelitian ini
bertujuan untuk enganalisis efek pemberian TGF-β1 rekombinan manusia pada sel
punca kanker payudara (aldehyde dehydrogenase positive, ALDH+) terhadap
ekspresi marker kepuncaan melalui pensinyalan autokrin TGF-β1. Sebagai
pembanding digunakan kanker payudara subtipe triple negative (TNBC).
Metode: BCSCs manusia (ALDH+) dan TNBC (MDA-MB-231) dikultur dalam
Dulbecco's Modified Eagle Medium/Nutrient Mixture F12/HG (DMEM F12/HG)
dengan suplemen 0,1 ng/ml protein rekombinan TGF-β1 manusia (rhTGF-β1)
selama periode 1, 2 dan 4 jam. Medium kultur kemudian diganti dengan DMEM
F12/HG tanpa serum selama 24 jam. Tingkat ekspresi mRNA reseptor TGF-β tipe
1 (TβR1), TGF-β1, faktor transkripsi pengikat oktamer 4 (OCT4), dan anggota A1
keluarga aldehida dehidrogenase 1 (ALDH1A1) dianalisis menggunakan real time
reverse transcriptase polymerase chain reactions (RT-qPCR). Kadar protein TGF-
β1 dalam media kultur ditentukan dengan menggunakan enzyme-linked
immunosorbent assay (ELISA). Sifat tumorigenik sel diuji dengan uji
mammosphere forming unit (MFU assay).
Hasil: Tingkat ekspresi mRNA dan protein TGF-β1 BCSC setelah perlakuan
tampak meningkat namun tidak pada TNBC. mRNA TβR1 BCSCs meningkat pada
periode perlakuan 1 dan 2 jam, sedangkan pada TNBC hanya pada periode 1 jam.
Penanda kepuncaan ALDH1A1 dan OCT4 tampak meningkat pada BCSC namun
tidak pada TNBC. Uji MFU menunjukkan sifat tumorigenik kedua kelompok sel
terutama pada periode perlakuan 2 jam tampak meningkat.
Kesimpulan: Perlakuan TGF-β1 dalam konsentrasi rendah dan dalam waktu
singkat memicu autoinduksi pada BCSCs yang menyebabkan peningkatan ekspresi
gen kepuncaan melalui pensinyalan autokrin. Sedangkan pada TNBC, peningkatan
ekspresi marker kepuncaan tidak terjadi. Namun demikian, sifat tumorigenik BCSC
dan TNBC tetap meningkat.

Background: Breast Cancer Stem Cells (BCSC) is a population of breast cancer
cells that have stem cell characteristics. BCSCs maintain tumor stability by
initiating the formation of new cancer cell populations and providing resistance to
therapy. BCSCs can interact with the tumor microenvironment which releasing
various cytokines and growth factors, including TGF-β1. Through the
autoinduction mechanism, TGF-β1 can increase endogenous TGF-β1 production
and autocrine signaling. TGF-β1 autocrine signaling can increase the tumor
promoter role of TGF-β1 in cancer development by enhancing the stemness and
tumorigenic properties. This study aims to analyze the effect of Human TGF-β1
recombinant protein treatment to breast cancer stem cells (aldehyde dehydrogenase
positive, ALDH+) on the expression of stemness marker through TGF-β1 autocrine
signaling. Triple negative breast cancer (TNBC) was used as a comparison.
Methods: Human BCSCs (ALDH+) and TNBC (MDA-MB-231) were cultured in
Dulbecco's Modified Eagle Medium/Nutrient Mixture F12/HG (DMEM F12/HG)
with 0.1 ng / ml recombinant protein of human TGF-β1 supplementation (rhTGF-
β1) over 1, 2 and 4 hour periods. The culture medium was then replaced with
DMEM F12/HG serum-free for 24 hours. The expression levels of the TGF-β
receptor type 1 (TβR1), TGF-β1, octamer-binding transcription factor 4 (OCT4),
and members of the A1 family of aldehyde dehydrogenase 1 (ALDH1A1) mRNA
were analyzed using real time reverse transcriptase polymerase chain reactions
(RT-qPCR). TGF-β1 protein levels in conditioned medium were determined using
an enzyme-linked immunosorbent assay (ELISA). The tumorigenic properties of
cells were tested by the mammosphere forming unit (MFU) assay.
Results: The expression level of mRNA and TGF-β1 BCSC protein after treatment
appeared to be increased but not in TNBC. mRNA TβR1 BCSCs increased in the
treatment period of 1 and 2 hours, whereas in TNBC only in the 1 hour period. The
markers of ALDH1A1 and OCT4 expression appeared to be increased in BCSC but
not in TNBC. The MFU test showed that the tumorigenic properties of both cell
groups, especially in the 2 hour treatment period, appeared to be increasing.
Conclusion: Treatment of TGF-β1 in low concentrations and in a short time
triggered autoinduction of BCSCs and leads to the increased expression of stemness
genes marker via autocrine signaling. Whereas in TNBC, this increase in the
expression of the stemness markers did not occur. However, the tumorigenic nature
of BCSC and TNBC continues to increase.

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Jenis Koleksi : UI - Tesis Membership
No. Panggil : T-pdf
Entri utama-Nama orang :
Entri tambahan-Nama orang :
Entri tambahan-Nama badan :
Program Studi :
Subjek :
Penerbitan : Jakarta: Fakultas Kedokteran Universitas Indonesia, 2020
Bahasa : ind
Sumber Pengatalogan : LibUI ind rda
Tipe Konten : text
Tipe Media : computer
Tipe Carrier : online resource
Deskripsi Fisik : xiv, 69 pages : illustration + appendix
Naskah Ringkas :
Lembaga Pemilik : Universitas Indonesia
Lokasi : Perpustakaan UI
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