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標題: | 探討UDP-半乳糖轉運蛋白SLC35A2在神經母細胞瘤中的角色 To investigate the role of UDP-galactose transporter SLC35A2 in Neuroblastoma |
作者: | 張秀羚 Hsiu-Ling Chang |
指導教授: | 林能裕 Neng-Yu Lin |
關鍵字: | 神經母細胞瘤,UDP-半乳糖轉運蛋白(UGT),SLC35A2,自噬,細胞凋亡, Neuroblastoma,UDP-galactose transporter (UGT),SLC35A2,autophagy,apoptosis, |
出版年 : | 2023 |
學位: | 碩士 |
摘要: | 神經母細胞瘤(NB)是兒童期常見的惡性實體瘤,起源於神經母細胞的異常分化,主要影響交感神經節和腎上腺髓質。它約佔兒童癌症相關死亡的15%。
細胞表面的異常醣基化為癌細胞的顯著特徵之一,與惡性細胞轉化有關。這種非典型醣基化可能是由於生物合成酶的異常所造成,例如糖基轉移酶和核苷酸糖轉運蛋白。溶質載體家族35成員A2(SLC35A2),為UDP-半乳糖轉運蛋白(UGT),會將UDP-半乳糖轉運至高基氏體囊泡中參與醣基化。SLC35A2突變與先天性醣基化障礙疾病相關,但其在神經母細胞瘤中的作用仍不清楚。 在本研究中,我們透過臨床病理、生物學因素及免疫組織化學確認SLC35A2和MYCN在神經母細胞瘤中的臨床相關性。首先我們使用定量實時PCR分析了14種人類NB細胞系中SLC35A2的表現量。隨後,我們敲落或過表現SLC35A2基因以研究其對GI-ME-N、SKN-BE和SH-SY5Y細胞系表型的影響。研究結果表明,SLC35A2過表現顯著降低GI-ME-N的細胞活力,而在SKN-BE及SH-SY5Y細胞中抑制SLC35A2表現都顯著增加細胞活力。為了評估醣基化變化,我們使用了識別GlcNAc的LEL和STL等凝集素,結果顯示與對照組相比,SKN-BE細胞中的SLC35A2敲落導致GlcNAcylation增加。 此外,本研究還探討了SLC35A2在神經母細胞瘤中自噬作用和細胞凋亡的潛在參與。由於細胞自噬及細胞凋亡失調在多種疾病包括癌症有關。利用細胞凋亡測定顯示,與對照組相比,在葡萄糖剝奪條件下,SLC35A2敲落的SKN-BE細胞較不易死亡,而在穿透式電子顯微鏡及西方墨點法結果顯示,SLC35A2敲落會促進SKN-BE細胞自噬。總之,本研究強調了SLC35A2在調節神經母細胞瘤中醣蛋白改變和葡萄糖代謝中的潛在作用,並表明自噬的參與。然而,還需要進一步的研究來闡明SLC35A2和自噬作用在神經母細胞瘤發病機制中的潛在機制和影響。 Neuroblastoma (NB) is a common malignant solid tumor in childhood that originates from abnormal differentiation of neuroblasts, primarily affecting the sympathetic ganglia and adrenal medulla. It accounts for approximately 15% of childhood cancer-related deaths. Aberrant glycosylation on the cell surface is one of the prominent characteristics of cancer cells and is associated with malignant cell transformation. This atypical glycosylation may be caused by abnormalities in biosynthetic enzymes such as glycosyltransferases and nucleotide sugar transporters. Solute carrier family 35 member A2 (SLC35A2), also known as UDP-galactose transporter (UGT), transports UDP-galactose into the Golgi apparatus vesicles for glycosylation. Mutations in SLC35A2 are associated with congenital disorders of glycosylation, but its role in neuroblastoma remains unclear. In this study, we investigated the clinical relevance of SLC35A2 and MYCN in neuroblastoma through clinical pathology, biological factors, and immunohistochemistry. Firstly, we analyzed the expression levels of SLC35A2 in 14 human neuroblastoma cell lines using quantitative real-time PCR. Subsequently, we knocked down or overexpressed the SLC35A2 gene to study its effects on the phenotype of GI-ME-N, SKN-BE, and SH-SY5Y cell lines. The results showed that overexpression of SLC35A2 significantly reduced cell viability in GI-ME-N cells, while inhibiting SLC35A2 expression in SKN-BE and SH-SY5Y cells significantly increased cell viability. To evaluate glycosylation changes, we used lectins such as LEL and STL that recognize GlcNAc residues, and the results showed that SLC35A2 knockdown in SKN-BE cells led to increased GlcNAcylation compared to the control group. Furthermore, this study also investigated the potential involvement of SLC35A2 in autophagy and apoptosis in neuroblastoma. Dysregulation of autophagy and apoptosis is associated with various diseases, including cancer. Apoptosis assays demonstrated that SKN-BE cells with SLC35A2 knockdown were less prone to cell death under glucose deprivation conditions, and transmission electron microscopy and Western blotting results indicated that SLC35A2 knockdown promoted autophagy in SKN-BE cells. In conclusion, this study highlights the potential role of SLC35A2 in regulating glycoprotein alterations and glucose metabolism in neuroblastoma and suggests the involvement of autophagy. However, further research is needed to elucidate the potential mechanisms and impacts of SLC35A2 and autophagy in the pathogenesis of neuroblastoma. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/90830 |
DOI: | 10.6342/NTU202303651 |
全文授權: | 同意授權(限校園內公開) |
顯示於系所單位: | 解剖學暨細胞生物學科所 |
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