Analysis of pluripotency marker expression in human glioblastoma multiforme cells treated with conditioned medium of umbilical cord-derived mesenchymal stem cells

Hardiany, Novi Silvia; Purnamawati,; Dewi, Syarifah; Paramita, Reni; Wanandi, Septelia Inawati

doi:10.12688/f1000research.13154.1

Cited by 6 publications

(6 citation statements)

References 21 publications

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“…Our experimental setup was similar to that proposed by Hardiany et al (14). About 400,000 T98G cells were plated in triplicate in 12-well plates in high-glucose DMEM with 10% FBS, 1% amphotericin B and 1% streptomycin-penicillin and were incubated overnight.…”

Section: Methodsmentioning

confidence: 82%

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Gene Expression of Molecules Regulating Apoptotic Pathways in Glioblastoma Multiforme Treated with Umbilical Cord Stem Cell Conditioned Medium

Hardiany

Ngadiono

et al. 2019

MJMS

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BackgroundGlioblastoma multiforme (GBM) is the most malignant primary brain tumour and there is no definite cure. It has been suggested that there are significant interactions among mesenchymal stem cells (MSCs), their released factors and tumour cells that ultimately determine GBM’s growth pattern. This study aims to analyse the expression of molecules involved in GBM cell apoptotic pathways following treatment with the MSC secretome.MethodsA conditioned medium of umbilical cord-derived MSCs (UCMSC-CM) was generated by culturing the cells on serum-free αMEM for 24 h. Following this, human GBM T98G cells were treated with UCMSC-CM for 24 h. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was then performed to measure the mRNA expression of survivin, caspase-9, TNF-related apoptosis-inducing ligand (TRAIL), DR4 and DcR1.ResultsmRNA expression of caspase-9 in CM-treated T98G cells increased 1.6-fold (P = 0.017), whereas mRNA expression of survivin increased 3.5-fold (P = 0.002). On the other hand, TRAIL protein expression was upregulated (1.2-fold), whereas mRNA expression was downregulated (0.4-fold), in CM-treated cells. Moreover, there was an increase in the mRNA expression of both DR4 (3.5-fold) and DcR1 (1,368.5-fold) in CM-treated cells.ConclusionThe UCMSC-CM was able to regulate the expression of molecules involved in GBM cell apoptotic pathways. However, the expression of anti-apoptotic molecules was more upregulated than that of pro-apoptotic molecules.

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Section: Methodsmentioning

confidence: 82%

“…Thereafter, the samples were centrifuged to remove any cell debris and passed through a 0.22 μm filter. The resulting UCMSC-CM was then diluted with fresh high-glucose DMEM to achieve a final concentration of 50% (14).…”

Section: Methodsmentioning

confidence: 99%

Gene Expression of Molecules Regulating Apoptotic Pathways in Glioblastoma Multiforme Treated with Umbilical Cord Stem Cell Conditioned Medium

Hardiany

Ngadiono

et al. 2019

MJMS

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“…(10) Our previous study conclude that treated T98G cells with CM of MSCs could rise pluripotency marker such as Sex determining region Y box 2 (SOX2), Octamer binding transcription factor 4 (OCT-4), and homeobox Nanog protein. (11) In addition, our another study proved that CM-UCSCs does not seems affect apoptosis but tend to increase cell proliferation in T98G cells. (12) Nevertheless, signaling cascade pathway in GBM cells treated with CM of umbilical cord-derived mesenchymal stem cells (UCMSC) is not well established.…”

Section: Introductionmentioning

confidence: 80%

“…UCSCs were provided from our preious research (11) which were grown in a Gibco α-Minimum Essential Medium (α-MEM) (Thermo Fisher Scientific) on a 25 cm 2 T-Flask with 10 % fetal bovine serum, 1% penicillin-streptomycin, and 1% amphotericin. Culture cells were placed in chamber with 37 o C, 5% saturation CO 2 , and 95% saturation O 2 .…”

Section: Culture Of Ucscs and Preparation Of CMmentioning

confidence: 99%

TNFR, TRAF2, NF-κB mRNA Levels of Glioblastoma Multiforme Cells Treated by Conditioned Medium of Umbilical Cord-derived Mesenchymal Stem Cells

2019

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BACKGROUND: Glioblastoma multiforme (GBM) is a human malignant brain tumor which is arise from glial cells. Our previous study proved that GBM cells proliferation increased after treating by conditioned medium of umbilical cord-derived mesenchymal stem cells (CM-UCSCs). Cells proliferation is probably mediated by tumor necrosis factor (TNF)-α which could bind to membrane receptor and induce signaling pathway. Therefore, this research was intended to analyze the mRNA expression of TNF-α signaling pathway molecules on CM-treated GBM cells by measuring TNF receptor 1 and 2 (TNFR1 and TNFR2), TNFR associated factor 2 (TRAF2), nuclear factor kappa B (NF-κB) mRNA level, and TNFR2 protein level.METHODS: UCSCs and human glioblastoma T98G cells were cultured and harvested after 80% confluence. CM was prepared by growing UCSCs in serum alpha Minimum Essential Media (α-MEM) for 24 hours. Fifty percent concentration of CM-UCSCs was used to treat T98G cells for 24 hours. TNF-α level in CM-UCSC was detected using enzyme linked-immunosorbent assay (ELISA), while the expression of TNFR1, TNFR2, TRAF2 and NF-κB were detected using quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR), and TNFR2 protein level was detected using sandwich ELISA.RESULTS: TNF-α level was detected in CM-UCSCs 4.4 pg/mL. Moreover, the expression of TNFR1, TNFR2, TRAF2 and NF-κB were significantly 1.4-fold, 4.9-fold, 5.6-fold, 1.8-fold respectively higher in T98G treated cells than control. TNFR2 protein level in T98G treated cells was 11.57 pg/mg protein higher than control.CONCLUSION: The expression of molecules involved in TNF-α signaling pathway were up regulated in T98G cells treated by CM-UCSCs.KEYWORDS: CM-UCSCs, TNFR1, TNFR2, TRAF2, NF-κB, T98G cells

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“…(9) Selain penelitian di atas, penggunaan sekretom sel punca mesenkim mampu meningkatkan ekspresi mRNA SOX2 dan OCT4 pada sel glioblastoma T98G. (10) Lebih jauh juga dibuktikan bahwa penggunaan sekretom sel punca mesenkim adiposa dan umbilical meningkatkan ekspresi mRNA ALDH1A1 dan ALDH1A3 pada sel kanker payudara manusia. (11) Beberapa petanda dapat diajukan sebagai indikator kepuncaan diantaranya CD133, SOX2, Nestin, dan A2B5.…”

Section: Abstract Pendahuluanunclassified

Pendeteksian petanda kepuncaan glioblastoma multiforme

Yohana¹

2020

J Biomedika Kesehat

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Glioblastoma multiforme (GBM) adalah tumor otak ganas yang memiliki populasi sel punca kanker yang dapat mempertahankan formasi tumor. Peranan sel punca kanker telah banyak dipelajari yang memiliki tanggung jawab terhadap resistensi dan rekurensi terapi GBM seperti radiasi dan kemoterapi. Beberapa petanda kepuncaan dapat dipakai diantaranya CD133, Nestin, A2B5, CD44, SOX2 and OCT4. Pemeriksaan histopatologi terhadap jaringan tumor yang dioperasi menjadi standar baku untuk menentukan derajat, tingkat keganasan, dan prognosis keganasan. Namun di sisi lain, kehadiran populasi sel punca yang memiliki sifat mampu memperbaharui diri dan mampu menginduksi pembentukan tumor memerlukan pemeriksaan yang lebih mendalam mengenai karakteristik biologi sel tumor. Pemeriksaan sel punca dilakukan menggunakan flowcytometry dan imunohistokimia. Pemeriksaan petanda kepuncaan glioblastoma dilakukan dengan quantitative Reverse Transcriptase Real Time Polymerase Chain Reaction (qRT-PCR), Enzyme Linked Immunoabsorbent Assay (ELISA), Western Blot, Imunohistokimia dan flowcytometry. Petanda CD133 ditemukan ekspresinya meningkat pada berbagai pemeriksaan. CD133 digunakan sebagai prognostik GBM.

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Analysis of pluripotency marker expression in human glioblastoma multiforme cells treated with conditioned medium of umbilical cord-derived mesenchymal stem cells

Cited by 6 publications

References 21 publications

Gene Expression of Molecules Regulating Apoptotic Pathways in Glioblastoma Multiforme Treated with Umbilical Cord Stem Cell Conditioned Medium

Gene Expression of Molecules Regulating Apoptotic Pathways in Glioblastoma Multiforme Treated with Umbilical Cord Stem Cell Conditioned Medium

TNFR, TRAF2, NF-κB mRNA Levels of Glioblastoma Multiforme Cells Treated by Conditioned Medium of Umbilical Cord-derived Mesenchymal Stem Cells

Pendeteksian petanda kepuncaan glioblastoma multiforme

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