TY  - JOUR
T1  - Toll-like receptor 3-mediated modulation of umbilical cord mesenchymal stem cell phenotype and pancreatic cancer cell responses during coculture
AU  - Kaçaroğlu, Demet
AU  - Yılmaz, Ayşegül
PY  - 2025
DA  - October
Y2  - 2025
JF  - Trakya University Journal of Natural Sciences
JO  - Trakya Univ J Nat Sci
PB  - Trakya University
WT  - DergiPark
SN  - 2528-9691
SP  - 213
EP  - 222
VL  - 26
IS  - 2
LA  - en
AB  - Mesenchymal stem cells (MSCs) are progenitor cells isolated from various tissues and are crucial for tissue repair, immune support, and anticancer therapies. MSC functions such as migration, immunomodulation, and regeneration are regulated through Toll-like receptors (TLRs). In particular, TLR3 activation enhances the immunosuppressive and therapeutic capabilities of MSCs. This research employed human umbilical cord-derived MSCs (UCMSCs) and investigated the effects of TLR3 stimulation on their viability, phenotype-associated gene expression, and during co-culture with Panc-1 pancreatic cancer cells. UCMSCs were cultured and characterized for mesenchymal markers by flow cytometry. TLR3- based signaling was modulated using Poly(A:U) (an agonist) and CU-CPT4a (an antagonist). Cell viability was assessed using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, and relative gene expression was measured employing quantitative reverse transcription polymerase chain reaction. Panc-1 cells were co-cultured with UCMSCs to evaluate TLR3-mediated effects. Data are presented as the means ± standard error of the mean, with statistical significance determined by analysis of variance (p ≤ 0.05). The TLR3 agonist improved cell viability, whereas the antagonist reduced it. Additionally, both regulated the expression of CD44, CDH1, and VIMs. When UCMSCs and Panc-1 cells were cocultured at 10:1, TLR3 affected the expression of MSC-related genes, including CD44, CDH1, CLDN1, VIM, ZEB1, MMP9, MMP2, TIMP1, VEGFR2, and PLAU. Thus, TLR3-based signaling influenced the viability, maintenance of the mesenchymal phenotype, and Panc-1 coculture-associated phenotype in UCMSCs. These results underscore the crucial role of TLR3-based signaling in modulating UCMSC function and suggest its potential utility in enhancing MSC-based therapeutic strategies. We believe that these results can help elucidate the role of TLR3-based signaling on UCMSC functions and provide a basis for future research.
KW  - Mesenchymal stem cell
KW  - TLR3
KW  - UCMSC
KW  - pancreatic ductal adenocarcinoma
KW  - cancer biology
N2  - Mezenkimal kök hücreler (MSC’ler), doku onarımı, bağışıklık desteği ve kanser karşıtı tedaviler açısından kritik öneme sahip progenitör hücrelerdir. Farklı dokulardan izole edilen MSC’lerin göç, immünomodülasyon ve rejenerasyon gibi fonksiyonları Toll-benzeri reseptörler (TLR’ler) aracılığıyla düzenlenmektedir. Özellikle, TLR3 aktivasyonu MSC’lerin immünosupresif ve terapötik özelliklerini artırmaktadır. Bu çalışmada, TLR3 uyarımının insan göbek kordonu kaynaklı MSC’lerin (UK-MSC’ler) canlılığı, fenotip ile ilişkili gen ekspresyonu üzerindeki etkileri ile bu hücrelerin Panc-1 pankreas kanseri hücreleriyle ortak kültürdeki etkileri araştırılmıştır. UK-MSC’ler kültür ortamında çoğaltıldı ve mezenkimal yüzey belirteçleri açısından akım sitometrisi ile karakterize edildi. TLR3 sinyali, Poly(A:U) agonisti ve CU-CPT4a antagonisti ile modüle edildi. Hücre canlılığı 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolyum bromür testi ile değerlendirildi ve gen ekspresyonu gliseraldehit- 3-fosfat dehidrogenaz referans geni kullanılarak kantitatif gerçek zamanlı ters transkripsiyon polimeraz zincir reaksiyonu ile ölçüldü. Panc-1 pankreas kanseri hücreleri UK-MSC’lerle ko-kültüre edilerek TLR3 aracılı etkiler analiz edildi. Tüm veriler ortalama ± ortalamanın standart hatası olarak sunuldu ve istatistiksel analiz varyans analizi veya t-test ile yapıldı (p ≤ 0,05). TLR3 agonistlerinin hücre canlılığını artırdığı, TLR3 antagonistlerinin ise azalttığı gözlemlenmiştir. Ayrıca, hem agonist hem de antagonistlerin CD44, CDH1 ve VIM genlerinin ekspresyonunu düzenlediği bulunmuştur. UK-MSC ve Panc-1 hücreleri 10:1 oranında ortak kültüre alındığında, TLR3 aktivasyonu ve inhibisyonunun CD44, CDH1, CLDN1, VIM, ZEB1, MMP9, MMP2, TIMP1, VEGFR2 ve PLAU gibi MSC ile ilişkili genlerin ekspresyon profillerini etkilediği gösterilmiştir. Sonuç olarak, TLR3 sinyal iletiminin UK-MSC’lerin canlılığını, mezenkimal fenotipin korunmasını ve Panc-1 kanser hücrelerine yanıtla ilişkili fenotipini etkilediği gözlemlenmiştir. Bu bulgular, TLR3 sinyal iletiminin UK-MSC fonksiyonlarını modüle etmedeki önemini vurgulamakta ve özellikle tümörle ilişkili uygulamalarda MSC tabanlı tedavi stratejilerini geliştirmede potansiyel bir araç olabileceğini düşündürmektedir. Bu çalışmanın, TLR3 sinyal iletiminin UK-MSC’lerin biyolojik işlevleri üzerindeki etkilerini aydınlatmaya yardımcı olacağına ve gelecekteki araştırmalara temel oluşturacağına inanmaktayız.
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UR  - https://dergipark.org.tr/en/pub/trkjnat/issue//1709831
L1  - https://dergipark.org.tr/en/download/article-file/4917564
ER  -