Glioblastoma hücre hattında (U87) siklopamin ve temozolomid kombine tedavisinin miR-20a ekspresyonu üzerine etkileri

Year 2021, , 1426 - 1432, 30.12.2021

Leman Sencar , Derviş Mansuri Yılmaz Dilek Göktürk , Sema Özandaç Polat , Gülfidan Coşkun , Dilek Şaker , Tuğçe Sapmaz , Samet Kara Alper Çelenk , Sait Polat

https://doi.org/10.17826/cumj.996520

Abstract

Amaç: Bu çalışmada, glioblastoma hücre hattında (U87) siklopamin ve temozolomid (TMZ) tedavisinin ayrı ayrı ve kombine olarak uygulanmasından sonra miR-20a ekspresyonunun araştırılması ve bu miRNA’nın glioblastoma tedavisindeki etkinliğinin değerlendirilmesi amaçlanmıştır.
Gereç ve Yöntem: U87 MG, tedaviden önce 35 mm'lik kültür kabı üzerine yerleştirildi ve çoğaltıldı. Çalışma kapsamında 7 grup oluşturuldu: Grup 1: Kontrol grubu, Grup 2: Sham grubu (Dimetil sülfoksit), Grup 3: 2 Gün TMZ, Grup 4: 5 Gün TMZ, Grup 5: 3 saat siklopamin, Grup 6: 2 Gün TMZ + 3 saat siklopamin, Grup 7: 5 Gün TMZ + 3 saat siklopamin. Tedaviden sonra total miRNA izolasyonu ve qRT-PCR yapıldı.
Bulgular: Glioblastoma hücre hattında miR-20a ekspresyonunun grup 3, grup 4, grup 5, grup 6 ve grup 7’de belirgin olarak azaldığı tespit edildi. Ancak bu azalmanın en fazla grup 7 de olduğu saptandı . Ek olarak grup 7’de hücre sayısının azaldığı görüldü.
Sonuç: Glioblastoma hücrelerinde miR-20a’nın yüksek oranda eksprese olduğu; ancak siklopamin ve TMZ tedavilerinden sonra ekspresyonun azaldığı tespit edildi. miR-20a’nın glioblastoma için yeni bir terapötik hedef ve biyobelirteç olarak kullanılabileceği, siklopamin ve TMZ’nin glioblastomada tedavi amaçlı kullanılabileceği ancak bu konuda daha ileri çalışmaların yapılması gerektiği sonucuna varıldı.

Keywords

Glioblastoma, siklopamin, temozolomid, MiR-20a.

Supporting Institution

ÇUKUROVA ÜNİVERSİTESİ BİLİMSEL ARAŞTIRMA PROJELERİ

Project Number

TSA-2018-11296

Effects of cyclopamine and temozolomide combined treatment on miR-20a expression in glioblastoma cell line (U87)

Abstract

Purpose: The aim of this study was to investigate miR-20a expression in glioblastoma cell line (U87) after applying cyclopamine and temozolomide (TMZ) treatment separately and in combination and to evaluate the efficiency of miR-20a in glioblastoma treatment.
Materials and Methods: U87 MG was placed on a 35 mm culture dish before treatment. Within the scope of the study, 7 groups were formed: Group 1: Control group, Group 2: Sham group (Dimethyl sulfoxide), Group 3: 2 Days TMZ, Group 4: 5 Days TMZ, Group 5: 3 Hours Cyclopamine, Group 6: 2 Days TMZ + 3 Hours Cyclopamine, Group 7: 5 Days TMZ + 3 Hours Cyclopamine Total miRNA isolation and qRT-PCR was performed after treatment.
Results: MiR-20a expression in the glioblastoma cell line was found to be significantly decreased in group 3, group 4, group 5, group 6 and group 7. However, it was determined that this decrease was highest in group 7. In addition, it was observed that the number of cells decreased in group 7.
Conclusion: MiR-20a is highly expressed in glioblastoma cells; however, it was determined that the expressions decreased after Cyclopamine and TMZ treatments. miR-20a can be used as a new therapeutic target and biomarker for glioblastoma, Cyclopamine and TMZ can be used for treatment in glioblastoma, but further studies are needed on this subject.

Keywords

Glioblastoma, cyclopamine, temozolomide, MiR-20a.

Project Number

TSA-2018-11296

References

• Demuth T, Berens ME. Molecular mechanisms of glioma cell migration and invasion. J Neurooncol. 2014;70:217–28.
• Adamson C, Kanu OO, Mehta AI, Di C, Lin N, Mattox AK et al. Glioblastoma multiforme: a review of where we have been and where we are going. Expert Opin Investig Drugs. 2009;18:1061-83.
• Conti A, Gulì C, La Torre D, Tomasello C, Angileri FF, Aguennouz M. Role of inflammation and oxidative stress mediators in gliomas. Cancers (Basel). 2010;26;2:693-712.
• Jiang C, Chen X, Alattar M, Wei J, Liu H. MicroRNAs in tumorigenesis, metastasis, diagnosis and prognosis of gastric cancer. Cancer Gene Ther. 2015;22:291-301.
• Zhang W, Dahlberg JE, TamW. MicroRNAs in tumorigenesis. Am J Pathol. 2007;171:728–38.
• Moller HG, Rasmussen AP, Andersen HH, Johnsen KB, Henriksen M, Duroux M. A systematic review of microRNA in glioblastoma multiforme: micro-modulators in the mesenchymal mode of migration and invasion. Mol Neurobiol. 2013;47:131-44.
• Hirose Y, Berger MS, Pieper RO. p53 Effects both the duration of G2/M arrest and the fate of temozolomide-treated human glioblastoma cells. Cancer Res. 2001;61:1957–63.
• Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352:987-96.
• Liu YJ, Ma YC, Zhang WJ, Yang ZZ, Liang DS, Wu ZF, Qi XR1. Combination therapy with micellarized cyclopamine and temozolomide attenuate glioblastoma growth through Gli1 down-regulation. Oncotarget. 2017;8:42495-509.
• Ulasov IV, Nandi S, Dey M, Sonabend AM, Lesniak MS. Inhibition of sonic hedgehog and notch pathways enhances sensitivity of CD133 glioma stem cells to temozolomide therapy. Mol Med. 2011;17:103-12.
• Balbous A, Renoux B, Cortes U, Milin S. Selective release of a cyclopamine glucuronide prodrug towards stem-like cancer cell inhibition in glioblastoma. Mol Cancer Ther. 2014;13:2159-69.
• Zhao Y, Cui X, Zhu W, Chen X, Shen C, Liu Z, Yang G, Liu Y, Zhao S. Synergistic regulatory effects of micrRNAs on brain glioma cells. Mol Med Rep. 2017;16:1409-16.
• Şaker D, Sencar L, Yılmaz DM, Polat S. Relationships between microRNA-20a and microRNA-125b expression and apoptosis and inflammation in experimental spinal cord injury. Neurol Res. 2019;41:991-1000.
• Bar EE, Chaudhry A, Lin A, Fan X. Cyclopamine-mediated hedgehog pathway inhibition depletes stem- like cancer cells in glioblastoma. Stem Cells. 2007;25:2524–33.
• Yang Y, Zhang X, Wang Y, Zhang, Gumi Z. miR-204 reverses temozolomide resistance and inhibits cancer initiating cells phenotypes by degrading FAP-α in glioblastoma. Oncol Lett. 2018; 15: 7563–70.
• Fang L, Wang X, Sun B, Zhang X. Expression, regulation and mechanism of action of miR-17-92 cluster in tumor cells. Int J Mol Med. 2017;40:1624–30.
• Zhang GJ, Li Y, Zhou H, Xiao HX, Zhou T. miR-20a is an independent prognostic factor in colorectal cancer and is involved in cell metastasis. Mol Med Rep. 2014;10:283-91.
• Cui C, Cui Y, Fu Y, Ma S, Zhang S. Microarray analysis reveals gene and microRNA signatures in diabetic kidney disease. Mol Med Rep. 2018;17:61–8.