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
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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