Meme kanserinde CLDN10'un hücre döngüsü, hücre proliferasyonu ve hücre ölümü üzerindeki etkilerinin incelenmesi
Küçük Resim Yok
Tarih
2024
Yazarlar
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Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Meme Kanserinde CLDN10'un Hücre Döngüsü, Hücre Proliferasyonu ve Hücre Ölümü Üzerindeki Etkilerinin İncelenmesi Amaç: Klaudinler, sıkı bağlantılarla ilişkili bir grup transmembran proteindir. Bu çalışmada klaudinlerden biri olan Klaudin-10 (CLDN10)'un karsinogenez sürecindeki rolünü araştırmak amaçlanmıştır. Bu amaçla çeşitli hücre hatlarında CLDN10'un ifadesi belirlenerek, CLDN10 geninin siRNA aracılığı ile baskılanması sonucunda hücre ölümü, proliferasyonu ve hücre döngüsündeki olası değişimler in vitro olarak araştırılmıştır. Materyal ve metot: Çalışmada ZR-751, BT-474, SKBR3, BxPC-3, MCF10A, MDA-MB-231 ve MCF7 hücre hatları kullanılmıştır. Bu hücrelerde CLDN10 geninin ifade edilme düzeyleri belirlenmiştir. Daha sonra CLDN10 proteinin meme kanseri hücrelerindeki varlığını belirlemek amacıyla Western Blot analizi yapılmıştır. CLDN10 geninin siRNA aracılığı ile baskılanması sonucu olası değişimleri belirlemek için MDA-MB-231 ve MCF7 hücre hatları kullanılmıştır. CLDN10'un baskılandığı hücre hatları ile kontrol gruplarında hücrelerin proliferasyon yetenekleri MTS testi, hücre döngüsü ve hücre ölümü bakımından farklılıkları flow sitometri ile analiz edilmiştir. İlave olarak koloni oluşturma ile invazyon bakımından da karşılaştırmalar yapılmıştır. Bulgular: CLDN10'un ifadesinin en fazla ZR-751 hücre hattında olduğu saptanmıştır. Bunu sırasıyla BxPC-3, MCF7, SKBR3, MCF10A, MDA-MB-231 hücre hatlarının takip ettiği gözlenmiştir. CLDN10 proteininin MDA-MB-231'de %60, MCF7'de %70, BT-474'de %83, ZR-751'de %83, SKBR3'de %61 oranında ifade edildiği belirlenmiştir. CLDN10 siRNA uygulaması sonucunda MCF7 ve MDA-MB-231 hücre hatlarında sırası ile proliferasyon %60 ve %57, koloni oluşturma yeteneği %42 ve %55, invazyon yeteneği ise %78 ve %42 oranında azalmıştır. CLDN10 siRNA uygulamasının hücre ölümünde bir farklılığa neden olmadığı bulunmuştur. Sonuç: Çalışılan tüm hücre hatlarında CLDN10'un çeşitli düzeylerde ifade edildiği belirlenmiştir. CLDN10'un ifadesinin baskılandığı MCF7 ve MDA-MB-231 hücrelerinde, proliferasyon, invazyon ve koloni oluşturma yeteneklerinin azaldığı görülmüştür. Elde edilen bulgular ile ilişkili olarak kanser tedavisinde CLDN10'un baskılanmasının yeni bir hedef olabileceği düşülmektedir. Anahtar Kelimeler: Meme Kanseri, CLDN10
Examination of The Effects of CLDN10 on Cell Cycle, Cell Proliferation and Cell Death in Breast Cancer Aim: Claudins are a group of transmembrane proteins associated with tight junctions. This study aimed to investigate the role of one of the claudins, Claudin-10 (CLDN10), in the carcinogenesis process. For this purpose, the expression of CLDN10 was determined in various cell lines and the possible changes in cell death, proliferation and cell cycle as a result of the suppression of the CLDN10 gene via siRNA were investigated in vitro. Material and Method: ZR-751, BT-474, SKBR3, BxPC-3, MCF10A, MDA-MB-231 and MCF7 cell lines were used in the study. The expression levels of CLDN10 gene were determined in these cells. Then, Western Blot analysis was performed to determine the presence of CLDN10 protein in breast cancer cells. MDA-MB-231 and MCF7 cell lines were used to determine possible changes as a result of suppression of CLDN10 gene via siRNA. Differences in proliferation abilities of cells in CLDN10 suppressed cell lines and control groups were analyzed by MTS test, cell cycle and cell death by flow cytometry. Additionally, comparisons were made in terms of colony formation and invasion. Results: t was determined that CLDN10 expression was highest in ZR-751 cell line. This was followed by BxPC-3, MCF7, SKBR3, MCF10A, MDA-MB-231 cell lines, respectively. It was determined that CLDN10 protein was expressed by 60% in MDA-MB-231, 70% in MCF7, 83% in BT-474, 83% in ZR-751, and 61% in SKBR3. As a result of CLDN10 siRNA application, proliferation decreased by 60% and 57%, colony formation ability by 42% and 55%, and invasion ability by 78% and 42% in MCF7 and MDA-MB-231 cell lines, respectively. It was found that CLDN10 siRNA application did not cause any difference in cell death. Conclusion: It was determined that CLDN10 was expressed at various levels in all studied cell lines. In MCF7 and MDA-MB-231 cells where CLDN10 expression was suppressed, proliferation, invasion and colony formation abilities were observed to be reduced. In relation to the findings obtained, it is thought that suppression of CLDN10 may be a new target in cancer treatment. Key Words: Breast Cancer, CLDN10
Examination of The Effects of CLDN10 on Cell Cycle, Cell Proliferation and Cell Death in Breast Cancer Aim: Claudins are a group of transmembrane proteins associated with tight junctions. This study aimed to investigate the role of one of the claudins, Claudin-10 (CLDN10), in the carcinogenesis process. For this purpose, the expression of CLDN10 was determined in various cell lines and the possible changes in cell death, proliferation and cell cycle as a result of the suppression of the CLDN10 gene via siRNA were investigated in vitro. Material and Method: ZR-751, BT-474, SKBR3, BxPC-3, MCF10A, MDA-MB-231 and MCF7 cell lines were used in the study. The expression levels of CLDN10 gene were determined in these cells. Then, Western Blot analysis was performed to determine the presence of CLDN10 protein in breast cancer cells. MDA-MB-231 and MCF7 cell lines were used to determine possible changes as a result of suppression of CLDN10 gene via siRNA. Differences in proliferation abilities of cells in CLDN10 suppressed cell lines and control groups were analyzed by MTS test, cell cycle and cell death by flow cytometry. Additionally, comparisons were made in terms of colony formation and invasion. Results: t was determined that CLDN10 expression was highest in ZR-751 cell line. This was followed by BxPC-3, MCF7, SKBR3, MCF10A, MDA-MB-231 cell lines, respectively. It was determined that CLDN10 protein was expressed by 60% in MDA-MB-231, 70% in MCF7, 83% in BT-474, 83% in ZR-751, and 61% in SKBR3. As a result of CLDN10 siRNA application, proliferation decreased by 60% and 57%, colony formation ability by 42% and 55%, and invasion ability by 78% and 42% in MCF7 and MDA-MB-231 cell lines, respectively. It was found that CLDN10 siRNA application did not cause any difference in cell death. Conclusion: It was determined that CLDN10 was expressed at various levels in all studied cell lines. In MCF7 and MDA-MB-231 cells where CLDN10 expression was suppressed, proliferation, invasion and colony formation abilities were observed to be reduced. In relation to the findings obtained, it is thought that suppression of CLDN10 may be a new target in cancer treatment. Key Words: Breast Cancer, CLDN10
Açıklama
Sağlık Bilimleri Enstitüsü, Tıbbi Biyoloji ve Genetik Ana Bilim Dalı
Anahtar Kelimeler
Tıbbi Biyoloji, Medical Biology
