SW1353, HEP3B ve HT-29 kanser hücre hatlarında ADAMTS9 gen ekspresyon analizi ve IL-23, IL-33, IL-6, IL-17F, IL-1 alfa sitokinleriyle uyarılma mekanizması
Küçük Resim Yok
Tarih
2013
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Turgut Özal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Bu çalışmada kondrosarkom, karaciğer kanseri ve kolorektal kansere ait hücre hatlarında ADAMTS9 gen ekspresyonunun bazı sitokinlerce 2, 6 ve 24. saatlerde olan değişimleri araştırılmıştır. Materyal-metod: Kondrosarkom (SW1353), karaciğer kanseri (HEP3B) ve kolon kanseri (HT-29) hücreleri IL-23, IL-33, IL-6, IL-17F ve IL-1alfa ile 2, 6 ve 24 saat muamele edildi. Trizolle RNA izolasyonu yapıldıktan sonra komplementer DNA?lar elde edildi. ADAMTS9 ve ß-Aktin çalışma sıcaklığını tespit etmek için gradient PCR yapıldı. Real-time PCR standartları için ADAMTS9 ve ß-Aktin PCRı kuruldu ve jel ekstraksiyonu yapıldı. Elde ettiğimiz DNA belirli oranlarda dilüe edildi. Uygun çalışma programında ADAMTS9 ve ß-Aktin real-time PCR?ları yapıldı. Bulgular: HEP3B hücre hattında IL-23 ve IL-33, ADAMTS9 ekspresyonunu süre bağımlı olarak istatistiksel olarak önemli oranda etkiledi (sırasıyla p=0.031 ve p=0.015). 6 saatlik IL-23 maruziyeti altında saptanan ADAMTS9 ekspresyonu, 2 saatlik maruziyete göre yüksek bulundu (p=0.029). 24 saatteki değeri ise 6 saat değerine göre önemli oranda düşüktü (p=0.029). IL-33 maruziyeti altında 24. saatteki ADAMTS9 ekspresyonu, hem 2 hem de 6 saatteki değerlerden düşük bulundu (p=0.029). HT29 hücre hattında ADAMTS9 ekspresyonu zamanla hiçbir sitokin maruziyeti altında önemli olarak artmadı. SW 1353 hücre hattında IL-6 maruziyeti altındaki 24. saat ADAMTS9 ekspresyonu, kontrol ve 6. saat değerine göre istatistiksel olarak düşük bulundu (p=0.029). Benzer şekilde kontrol grubundaki ADAMTS9 değeri, hem 2 saat hem de 24 saat IL-17F maruziyeti değerine göre sınırda bir önem düzeyinde yüksek bulundu (p=0.057). Sonuç: Karaciğer kanser hücrelerinde uzun dönemde düşük bulunan ADAMTS9 ekspresyonunun, IL-23 ve IL-33 maruziyeti ile kısa dönemde artmış olarak bulunması; kondrosarkomda ise IL-6 ve IL-17F ile ADAMTS9 ekspresyonunun sadece 6 saatlik dönemde artması, böylece tümöral dokuda düşük ekspresyonda saptanması beklenen ADAMTS9 düzeylerinin zaman bağımlı olarak sitokinlerce artırılması karşılıklı etkileşimlerin varlığını düşündürmekte ve daha ileri çalışmalarla ortaya konulması gerekmektedir.
Aim: The changes in ADAMTS9 gene expression in SW1353, HEP3B VE HT-29 cell lines with various cytokines at 2, 6 and 24 hours were investigated. Material and Methods: Condrosarcoma (SW1353), hepatic (HEP3B) and colorectal cancer (HT-29) cell lines were treated with IL-23, IL-33, IL-6, IL-17F ve IL-1alpha for about 2, 6 and 24 hours. Following RNA isolation with Trizol, complemantary DNAs were obtained, and gradient PCR was performed for determination of ß-Actin study temperature. ADAMTS9 and ß-Actin PCR were established and gel extraction was done for real-time PCR standarts. DNA obtained was diluted with a certain proportions. ADAMTS9 and ß-Actin real-time PCR were performed under the appropriate study program. Results: IL-23 ve IL-33 in the HEP3B cell line significantly affected ADAMTS9 expression in a time dependent manner (p=0.031 and p=0.015, respectively). ADAMTS9 expression found under 6-hour IL-23 exposure was significantly higher than under 2-hour (p=0.029). The 24-hour value was significantly lower than the 6- hour value (p=0.029). ADAMTS9 expression under IL-33 exposure at 24th hour was found as lower than the values at 2nd and 6th hour (p=0.029). ADAMTS9 expression in HT29 cell line did not significantly increased under cytokines herein during time period. 24-hour ADAMTS9 expression in SW1353 line under IL-6 exposure was statistically lower when compared to 6-hour value (p=0.029). Similarly ADAMTS9 value in control group was nearly higher than the values under 2- and 24-hour IL- 17F exposure (p=0.057). Conclusion: ADAMTS9 expression found as lower in hepatic cancer cells in long time period was increased under IL-23 and IL-33 exposure in short time period; ADAMTS9 expression with IL-6 and IL-17F was increased in only 6-hour period in chondrocarcinoma. Consequently increased ADAMTS9 expression, which would be expected as decreased in tumoral tissue otherwise, with cytokines as time dependent can be based on reciprocal interaction and therefore further studies should be done.
Aim: The changes in ADAMTS9 gene expression in SW1353, HEP3B VE HT-29 cell lines with various cytokines at 2, 6 and 24 hours were investigated. Material and Methods: Condrosarcoma (SW1353), hepatic (HEP3B) and colorectal cancer (HT-29) cell lines were treated with IL-23, IL-33, IL-6, IL-17F ve IL-1alpha for about 2, 6 and 24 hours. Following RNA isolation with Trizol, complemantary DNAs were obtained, and gradient PCR was performed for determination of ß-Actin study temperature. ADAMTS9 and ß-Actin PCR were established and gel extraction was done for real-time PCR standarts. DNA obtained was diluted with a certain proportions. ADAMTS9 and ß-Actin real-time PCR were performed under the appropriate study program. Results: IL-23 ve IL-33 in the HEP3B cell line significantly affected ADAMTS9 expression in a time dependent manner (p=0.031 and p=0.015, respectively). ADAMTS9 expression found under 6-hour IL-23 exposure was significantly higher than under 2-hour (p=0.029). The 24-hour value was significantly lower than the 6- hour value (p=0.029). ADAMTS9 expression under IL-33 exposure at 24th hour was found as lower than the values at 2nd and 6th hour (p=0.029). ADAMTS9 expression in HT29 cell line did not significantly increased under cytokines herein during time period. 24-hour ADAMTS9 expression in SW1353 line under IL-6 exposure was statistically lower when compared to 6-hour value (p=0.029). Similarly ADAMTS9 value in control group was nearly higher than the values under 2- and 24-hour IL- 17F exposure (p=0.057). Conclusion: ADAMTS9 expression found as lower in hepatic cancer cells in long time period was increased under IL-23 and IL-33 exposure in short time period; ADAMTS9 expression with IL-6 and IL-17F was increased in only 6-hour period in chondrocarcinoma. Consequently increased ADAMTS9 expression, which would be expected as decreased in tumoral tissue otherwise, with cytokines as time dependent can be based on reciprocal interaction and therefore further studies should be done.
Açıklama
Sağlık Bilimleri Enstitüsü, Tıbbi Genetik Ana Bilim Dalı
Anahtar Kelimeler
Genetik, Genetics
