Meme kanseri modeli olarak MCF-7 hücre hattında iyot uygulamasının hücre çoğalması üzerine etkisi
Küçük Resim Yok
Tarih
2024
Yazarlar
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Cilt Başlığı
Yayıncı
Trakya Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Giriş ve Amaç: Meme kanseri, günümüzde kadınlarda sıklığı ve ölüm oranlarıyla ilk sıradaki kanser türüdür. Cerrahi, radyoterapi, kemoterapi ve akıllı ilaç tedavileri uygulansa da halen alternatif tedavilere ihtiyaç vardır. Artan vakalar, koruyucu olduğu düşünülen takviye besinlerin, vitaminlerin, eser elementlerin ve birtakım bitkisel ürünlerin kullanımını arttırmıştır. İyotlu ürünler de bunlardan bazılarıdır. İyot kullanımı ve meme kanseri insidansındaki arasındaki bağlantı ilk olarak epidemiyolojik çalışmalarda fark edilmiş, ardından hayvan ve hücre çalışmaları ile desteklenmiştir. Çalışmamızda da, MCF-7 meme kanseri hücre hattında, iyot uygulamalarının çoğalma davranışına etkisini belirlemeyi, homeopatik amaçla alınan iyotlu ürünlerin, farkında olunmayan bir tümöre etkisini in vitroda simüle etmeyi amaçladık. Gereç ve Yöntemler: MCF-7 hücrelerinde değişen dozlarda (1, 2 ve 3 mM) iyot (I2) ve I2+KI uygulamalarıyla, çoğalma davranışı MTT, koloni formasyon, 3D sferoid kültür ve migrasyon deneyleri ile kalitatif olarak incelendi. Hücre çoğalma belirteci olan Ki-67 molekülünün immün floresan boyama ile pozitiflik oranları değerlendirildi. Bulgular: MTT deney sonuçlarına göre, en düşük dozda her iki iyot grubunda canlı hücre sayısında anlamlı bir azalma olmadığı, 2 mM dozda I2'nin, hücre canlılığında 48 saatte %20'lik bir düşüş sağladığı, I2+KI'nin canlı hücre sayısını 24 saatte %85'e, 48 saatte %74'e düşürdüğü belirlendi. En yüksek dozda 24 saatte I2, %61'e, I2+KI, %45'e, 48 saatte I2 %31'e, I2+KI %21'e düşürdü. Migrasyon deneyi ile, yüksek doz iyot gruplarında çizim kenarlarındaki yoğun düzensizleşmeler ölçümlerin sağlıklı yapılmasını engelledi. Analizler değerlendirme dışı bırakıldı. Koloni formasyon deneyi ile 3 mM I2'de canlı kalan hücreler gözlense de koloni oluşumu saptanmadı. Diğer gruplarda koloni formasyon belirlendi. I2+KI'da hücrelerin her üç dozda da kontrol grubuna oranla daha çok koloni oluşturduğu gözlendi. 3D kültürlerde I2'nin 2 mM ve 3 mM dozlarında, I2+KI'nin sadece 3 mM dozunda kontrole kıyasla sferoid alanında anlamlı bir artış görüldü. I2'nin 2 mM ve 3 mM dozlarında ve I2+KI'nın sadece 1 mM dozunda sferoid yoğunluğunda anlamlı değişiklik gözlendi. Ki-67 skorlaması, Ki-67 pozitif boyanan hücrelerin 3 mM I2+KI dışındaki gruplarda aritmetik olarak birbirine yakın olduğu, 3 mM I2+KI'de ise diğer gruplardan %28,5-%8,3 oranında daha düşük pozitiflik gözlenmiştir. Sonuç: I2 ve I2+KI uygulamasının MCF-7 hücrelerinde doza ve süreye bağlı olarak canlılığı azalttığı gösterilmiştir. Migrasyon deneyleri değerlendirme dışı bırakılmıştır. Farklı formlarda uygulanan iyot, koloni oluşturma kabiliyeti üzerine farklı etkiler göstermiştir. Etki, iyot formlarının farklı tutulumunun olabilmesine bağlanmıştır. 3D kültürlerde iyodun hücrelerarası bağlantıları zayıflattığı, bu durumun hücreyi apoptoza sürükleyebileceği ya da hücrenin epitelyal mezenkimal dönüşüm geçirebileceği şeklinde yorumlanmıştır. Ki-67 sağ kalan hücrelerde değerlendirildiğinden, iyot da ileri seviyede hücre kaybına yol açtığından, MCF-7 hücrelerinde, in vitroda Ki-67'nin verimli bir belirteç olmadığını düşünülmüştür. Anahtar kelimeler: MCF-7, Meme Kanseri, İyot, Ki-67, Proliferasyon
Introduction and Purpose: Breast cancer is the first cancer type in women today in terms of frequency and mortality rates. Although surgery, radiotherapy, chemotherapy and smart drug treatments are used, alternative treatmentsare still needed. Increasing cases have increased the use of nutritional supplements, vitamins, trace elements and some herbal products thought to be protective. Iodine products are some of them. The link between iodine use and breast cancer incidence was first recognized in epidemiological studies and subsequently supported by animal and cell studies. In our study, we aimed to determine the effect of iodine treatments on the proliferation behaviour of the MCF-7 breast cancer cell line and to simulate the effect of iodine products taken for homoeopathic purposes on an unknown tumour in vitro. Materials and Methods: Proliferation behavior was qualitatively examined by MTT, colony formation, 3D spheroid culture and migration experiments in MCF-7 cells with varying doses (1, 2 and 3 mM) of molecular iodine (I2) and I2+KI. Positivity rates of Ki-67 molecule, a cell proliferation marker, were evaluated with immunofluorescent staining. Results: According to the MTT experiment results, there was no significant decrease in the number of viable cells in both iodine groups at the lowest dose, I2 at a dose of 2 mM caused a 20% decrease in cell viability in 48 hours, and I2+KI increased the number of viable cells by 24 hours. It was determined that it decreased to 85% in an hour and to 74% in 48 hours. At the highest dose, I2 decreased to 61%, I2+KI to 45% in 24 hours, I2 to 31% and I2+KI to 21% in 48 hours. With the migration experiment, intense irregularities at the edges of the drawings in the high-dose iodine groups prevented accurate measurements. Analyzes were excluded from the evaluation. Although cells surviving in 3 mM I2 were observed with the colony formation assay, no colony formation was detected. Colony formation was determined in other groups. It was observed that the cells formed more colonies in I2+KI than in the control group at all three doses. In 3D cultures, a significant increase in spheroid area was observed at 2 mM and 3 mM doses of I2 and only at 3 mM dose of I2+KI compared to the control. A significant change in spheroid density was observed at 2 mM and 3 mM doses of I2 and only at 1 mM dose of I2+KI. Ki-67 scoring showed that Ki-67 positive stained cells were arithmetically close to each other in groups other than 3 mM I2+KI, while 28.5%-8.3% lower positivity was observed in 3 mM I2+KI than other groups. Conclusion: It has been shown that I2 and I2+KI treatments reduce the viability of MCF-7 cells depending on dose and time. Migration experiments were excluded from the evaluation. Iodine applied in different forms showed different effects on colony formation ability. The effect is attributed to the different retention of iodine forms. It has been interpreted that iodine weakens intercellular connections in 3D cultures, and this may lead the cell to apoptosis or undergo epithelial-mesenchymal transformation. Since Ki-67 was evaluated in surviving cells and iodine causes advanced cell loss, it was thought that Ki-67 was not a productive marker in vitro in MCF-7 cells. Key words: MCF-7, breast cancer, iodine, Ki-67, proliferation
Introduction and Purpose: Breast cancer is the first cancer type in women today in terms of frequency and mortality rates. Although surgery, radiotherapy, chemotherapy and smart drug treatments are used, alternative treatmentsare still needed. Increasing cases have increased the use of nutritional supplements, vitamins, trace elements and some herbal products thought to be protective. Iodine products are some of them. The link between iodine use and breast cancer incidence was first recognized in epidemiological studies and subsequently supported by animal and cell studies. In our study, we aimed to determine the effect of iodine treatments on the proliferation behaviour of the MCF-7 breast cancer cell line and to simulate the effect of iodine products taken for homoeopathic purposes on an unknown tumour in vitro. Materials and Methods: Proliferation behavior was qualitatively examined by MTT, colony formation, 3D spheroid culture and migration experiments in MCF-7 cells with varying doses (1, 2 and 3 mM) of molecular iodine (I2) and I2+KI. Positivity rates of Ki-67 molecule, a cell proliferation marker, were evaluated with immunofluorescent staining. Results: According to the MTT experiment results, there was no significant decrease in the number of viable cells in both iodine groups at the lowest dose, I2 at a dose of 2 mM caused a 20% decrease in cell viability in 48 hours, and I2+KI increased the number of viable cells by 24 hours. It was determined that it decreased to 85% in an hour and to 74% in 48 hours. At the highest dose, I2 decreased to 61%, I2+KI to 45% in 24 hours, I2 to 31% and I2+KI to 21% in 48 hours. With the migration experiment, intense irregularities at the edges of the drawings in the high-dose iodine groups prevented accurate measurements. Analyzes were excluded from the evaluation. Although cells surviving in 3 mM I2 were observed with the colony formation assay, no colony formation was detected. Colony formation was determined in other groups. It was observed that the cells formed more colonies in I2+KI than in the control group at all three doses. In 3D cultures, a significant increase in spheroid area was observed at 2 mM and 3 mM doses of I2 and only at 3 mM dose of I2+KI compared to the control. A significant change in spheroid density was observed at 2 mM and 3 mM doses of I2 and only at 1 mM dose of I2+KI. Ki-67 scoring showed that Ki-67 positive stained cells were arithmetically close to each other in groups other than 3 mM I2+KI, while 28.5%-8.3% lower positivity was observed in 3 mM I2+KI than other groups. Conclusion: It has been shown that I2 and I2+KI treatments reduce the viability of MCF-7 cells depending on dose and time. Migration experiments were excluded from the evaluation. Iodine applied in different forms showed different effects on colony formation ability. The effect is attributed to the different retention of iodine forms. It has been interpreted that iodine weakens intercellular connections in 3D cultures, and this may lead the cell to apoptosis or undergo epithelial-mesenchymal transformation. Since Ki-67 was evaluated in surviving cells and iodine causes advanced cell loss, it was thought that Ki-67 was not a productive marker in vitro in MCF-7 cells. Key words: MCF-7, breast cancer, iodine, Ki-67, proliferation
Açıklama
Yüksek Lisans
Anahtar Kelimeler
Tıbbi Biyoloji, Medical Biology
