Expression cloning of sialyltransferase involved in malignant progression of human RCC

唾液酸转移酶参与人肾细胞癌恶性进展的表达克隆

• 批准号: 09470338
• 负责人: SAITO Seiichi
• 金额: $ 8.45万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470338/
• 关键词: renal cell carcinoma; sialyltransterase; malignant progression; differentiation; globo-series ganglioside; 腎細胞癌; 転移; 発現クローニング; グロボシリーズ ガングリオシド

In all attempt to identify α 2, 3-sialyltransferase involved in synthesis of monosialosyl galactosyl globoside (MSGG), the first substance of globo-series ganglioside, associated with the malignant progression of human renal cell carcinoma (RCC), we utilized both expression cloning procedure and PCR method. Thus far, we could not obtain α 2, 3-sialyltransferase by expression cloning procedure, but 5' precursor of rRNA, placental transforming growth factor β, unknown cDNA, etc. were identified as concentrated cDNA by this procedure. We are now investigating the relationship between these cDNA and expression of MSGG.We obtained hST3Gal I and hST3Gal IV by PCR method. By Northern analysis, there was no clear correlation between hST3Gal I and the malignant progression of human RCC.None of the stable transfectants with hST3Gal I examined showed expression of MSGG.Stable transfectant with hST3Gal IV showed moderate increase of MSGG.However, hST3Gal IV transcript increased prominently in almost all normal renal tissues and decreased in most cases of cancer tissues. It was suggested that hST3Gal IV is associated with renal differentiation. From these results, it was anticipated that there is another a 2, 3-sialyltransferase determining the sysnthesis of MSGG.Investigation of this putative sialyltransferase is still underway. During this research, we identified N-acetyl-galactosaminyl disialosyl typel chain paragloboside (GalNAc DSPG-1) as a novel ganglioside, and found that DSGG is distributed in cells of mesoderm origin. We also found that expression of MSGG is inversely related to that of GM3 in human RCC.

为了鉴定与人肾细胞癌（RCC）恶性进展相关的单唾液酰半乳糖糖苷（MSGG）合成相关的α 2,3 -唾液酰转移酶（α 2,3 -唾液酰转移酶）是全球系列节脂苷的第一种物质。到目前为止，我们还不能通过表达克隆方法获得α 2,3 -唾液基转移酶，但通过该方法鉴定出rRNA的5'前体、胎盘转化生长因子β、未知cDNA等为浓缩cDNA。我们现在正在研究这些cDNA与MSGG表达之间的关系。采用PCR法获得hST3Gal I和hST3Gal IV。Northern分析发现hST3Gal I与人RCC的恶性进展没有明显的相关性。我检测的含有hST3Gal的稳定转染物均未显示MSGG的表达。稳定转染hST3Gal IV后，MSGG有中度升高。然而，hST3Gal IV转录本在几乎所有正常肾组织中显著升高，在大多数癌组织中显著降低。提示hST3Gal IV与肾分化有关。从这些结果可以推测，有另一种2,3 -唾液基转移酶决定了MSGG的合成。对这种假定的唾液转移酶的研究仍在进行中。在本研究中，我们鉴定了n -乙酰-半乳糖胺基二甲酰基型链副loboside （GalNAc DSPG-1）为一种新型神经节苷脂，并发现DSGG分布于中胚层起源的细胞中。我们还发现在人RCC中，MSGG的表达与GM3的表达呈负相关。