Expression of sialic acid binding receptors (siglecs) in human trophoblast cell line

Nazlı Çil; İ.veysel Fenkci; Gülçin Abban Mete; Doğukan Mutlu; Cihan Kabukçu; Ümit Çabuş

Article Details

Expression of sialic acid binding receptors (siglecs) in human trophoblast cell line

Authors:

Nazlı Çil | İ.veysel Fenkci | Gülçin Abban Mete | Doğukan Mutlu | Cihan Kabukçu | Ümit Çabuş

Publisher:

Pamukkale University

Main Topics:

Biyokimya ve Hücre Biyolojisi (Diğer)

Journal:

Pamukkale Tıp Dergisi Vol.17 Issue 2, pp.195-203

Document Type:

Article

Publication Date:

1/2024

Document Language:

English

Document ID (Org No.):

2001300671

pISSN:

1308-0865

Keywords:

Abstract:

 Purpose: Cell surface proteins known as Siglecs exhibit a specific affinity for sialic acid. Primarily located on the surface of immune cells, Siglecs belong to the subset of lectins called I-type lectins. Siglecs have important roles in maternal-fetal immune tolerance. We aimed to analyze the expression of Siglecs on Human Villous Trophoblasts HVT cells. Materials and methods: Total RNA was extracted from the HVT cell line, cDNA was synthesized, and real-time quantitative reverse transcription polymerase chain reaction RT-qPCR was performed the determine the changes in Siglec -3, -5, -6, -7, -8, -9, -10, -11, and -16 mRNA levels. In addition, Siglec levels were assessed by using immunohistochemical staining. Immunoreactivity against Siglec-6 and Siglec-9 was evaluated separately according to the intensity of brown color. Results: Expression levels of Siglec genes by qRT-PCR and melting curve analyses were performed using RNA extracted from the HVT cell line. Siglec -3, -5, -6, -7, -9, -10, -11, and -16 genes were found to be expressed in the HVT cell line. Differently, Siglec-8 results were undetected after cycle 40, which was considered a negative result. The immunocytochemical examination of the HVT cell line revealed that Siglec-6 expression was moderate in the cytoplasm Score:2. Siglec-9 expression was prominent in the cytoplasm Score:3. Conclusion: The results showed that Siglec-6 and Siglec-9 were expressed more than other Siglec proteins in the human trophoblast cells. Immunocytochemistry results also support these findings. Our study is the first to show Siglec-9 expression in human trophoblast cell.

 Purpose: Cell surface proteins known as Siglecs exhibit a specific affinity for sialic acid. Primarily located on the surface of immune cells, Siglecs belong to the subset of lectins called I-type lectins. Siglecs have important roles in maternal-fetal immune tolerance. We aimed to analyze the expression of Siglecs on Human Villous Trophoblasts HVT cells. Materials and methods: Total RNA was extracted from the HVT cell line, cDNA was synthesized, and real-time quantitative reverse transcription polymerase chain reaction RT-qPCR was performed the determine the changes in Siglec -3, -5, -6, -7, -8, -9, -10, -11, and -16 mRNA levels. In addition, Siglec levels were assessed by using immunohistochemical staining. Immunoreactivity against Siglec-6 and Siglec-9 was evaluated separately according to the intensity of brown color. Results: Expression levels of Siglec genes by qRT-PCR and melting curve analyses were performed using RNA extracted from the HVT cell line. Siglec -3, -5, -6, -7, -9, -10, -11, and -16 genes were found to be expressed in the HVT cell line. Differently, Siglec-8 results were undetected after cycle 40, which was considered a negative result. The immunocytochemical examination of the HVT cell line revealed that Siglec-6 expression was moderate in the cytoplasm Score:2. Siglec-9 expression was prominent in the cytoplasm Score:3. Conclusion: The results showed that Siglec-6 and Siglec-9 were expressed more than other Siglec proteins in the human trophoblast cells. Immunocytochemistry results also support these findings. Our study is the first to show Siglec-9 expression in human trophoblast cell.

 Purpose: Cell surface proteins known as Siglecs exhibit a specific affinity for sialic acid. Primarily located on the surface of immune cells, Siglecs belong to the subset of lectins called I-type lectins. Siglecs have important roles in maternal-fetal immune tolerance. We aimed to analyze the expression of Siglecs on Human Villous Trophoblasts HVT cells. Materials and methods: Total RNA was extracted from the HVT cell line, cDNA was synthesized, and real-time quantitative reverse transcription polymerase chain reaction RT-qPCR was performed the determine the changes in Siglec -3, -5, -6, -7, -8, -9, -10, -11, and -16 mRNA levels. In addition, Siglec levels were assessed by using immunohistochemical staining. Immunoreactivity against Siglec-6 and Siglec-9 was evaluated separately according to the intensity of brown color. Results: Expression levels of Siglec genes by qRT-PCR and melting curve analyses were performed using RNA extracted from the HVT cell line. Siglec -3, -5, -6, -7, -9, -10, -11, and -16 genes were found to be expressed in the HVT cell line. Differently, Siglec-8 results were undetected after cycle 40, which was considered a negative result. The immunocytochemical examination of the HVT cell line revealed that Siglec-6 expression was moderate in the cytoplasm Score:2. Siglec-9 expression was prominent in the cytoplasm Score:3. Conclusion: The results showed that Siglec-6 and Siglec-9 were expressed more than other Siglec proteins in the human trophoblast cells. Immunocytochemistry results also support these findings. Our study is the first to show Siglec-9 expression in human trophoblast cell.

 Amaç: Siglecler olarak bilinen hücre yüzeyi proteinleri, sialik asit için özel bir afinite sergilemektedir. Öncelikle bağışıklık hücrelerinin yüzeyinde bulunan Siglecler, I-tipi lektinler adı verilen lektin alt kümesine aittir. Siglecler maternal-fetal immün toleransta önemli rollere sahiptir. Bu çalışmada İnsan Villöz Trofoblast HVT hücre hatlarında Siglec tiplerinin belirlenmesi amaçlanmıştır. Gereç ve yöntem: HVT hücre hattından total RNA ekstrakte edildi, cDNA sentezlendi ve Siglec -3, -5, -6, -7, -8, -9, -10, -11 ve -16 gen ekspresyonu Gerçek Zamanlı Kantitatif Ters Transkripsiyon Polimeraz Zincir Reaksiyonu qRT-PCR ile analiz edildi. Ek olarak, Siglec seviyelerini belirlemek için immünohistokimyasal boyamalar yapılmıştır. Siglec-6 ve Siglec-9 a karşı immünoreaktivite kahverengi rengin yoğunluğuna göre ayrı ayrı değerlendirilmiştir. Bulgular: HVT hücre hattından ekstrakte edilen RNA kullanılarak Siglec genlerinin qRT-PCR ile ekspresyon seviyeleri ile erime eğrisi analizleri yapıldı. Siglec -3, -5, -6, -7, -9, -10, -11 ve -16 genlerinin HVT hücre hattında ifade edildiği bulundu. Farklı olarak, Siglec-8 sonuçları 40. döngüden sonra tespit edilemedi ve bu da negatif bir sonuç olarak kabul edildi. HVT hücre hattının immünositokimyasal incelemesinde Siglec-6 ekspresyonunun sitoplazmada orta düzeyde olduğu ortaya çıkmıştır Skor:2. Siglec-9 ekspresyonu sitoplazmada belirgindi Skor:3. Sonuç: Sonuçlar Siglec-6 ve Siglec-9un HVT hücre hattında diğer Siglec proteinlerinden daha fazla ifade edildiğini göstermiştir. İmmünositokimya sonuçları da bu bulguları desteklemektedir. Çalışmamız insan trofoblast hücrelerinde Siglec-9 ekspresyonunu gösteren ilk çalışmadır.

Article Details

Expression of sialic acid binding receptors (siglecs) in human trophoblast cell line

Authors:

Nazlı Çil | İ.veysel Fenkci | Gülçin Abban Mete | Doğukan Mutlu | Cihan Kabukçu | Ümit Çabuş

Publisher:

Pamukkale University

Main Topics:

Biyokimya ve Hücre Biyolojisi (Diğer)

Journal:

Pamukkale Tıp Dergisi Vol.17 Issue 2, pp.195-203

Document Type:

Article

Publication Date:

1/2024

Document Language:

English

Document ID (Org No.):

2001300671

pISSN:

1308-0865

Siglecs

human villous trophoblasts

sialic acidSiglecs

insan villöz trafoblast

sialik asit

Issues

Article Details

Expression of sialic acid binding receptors (siglecs) in human trophoblast cell line

Authors: Nazlı Çil | İ.veysel Fenkci | Gülçin Abban Mete | Doğukan Mutlu | Cihan Kabukçu | Ümit Çabuş

Publisher: Pamukkale University

Main Topics: Biyokimya ve Hücre Biyolojisi (Diğer)

Journal: Pamukkale Tıp Dergisi Vol.17 Issue 2, pp.195-203

Document Type: Article

Publication Date: 1/2024

Document Language: English

Document ID (Org No.): 2001300671

pISSN: 1308-0865

Siglecs

human villous trophoblasts

sialic acidSiglecs

insan villöz trafoblast

sialik asit

Issues

Authors:

Nazlı Çil | İ.veysel Fenkci | Gülçin Abban Mete | Doğukan Mutlu | Cihan Kabukçu | Ümit Çabuş

Publisher:

Pamukkale University

Main Topics:

Biyokimya ve Hücre Biyolojisi (Diğer)

Journal:

Pamukkale Tıp Dergisi Vol.17 Issue 2, pp.195-203

Document Type:

Article

Publication Date:

1/2024

Document Language:

English

Document ID (Org No.):

2001300671

pISSN:

1308-0865