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[细胞表面唾液酸及其连接方式对乳腺癌细胞黏附的影响]

[Effect of cell surface sialic acid and their linkages on adhesion of mammary carcinoma cells].

作者信息

Wang Xiao-yu, Lin Shao-qiang, Li Jun-wu, Kemmner Wolfgang, Ding Yan-qing

机构信息

Department of Obstetrics and Gynecology, First Affiliated Hospital of Ji'nan University, Guangzhou 510630, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2006 Jun;26(6):742-6.

Abstract

OBJECTIVE

To investigate the effect of cell surface sialic acid and its linkage on the cell-cell and cell-matrix adhesion of mammary carcinoma cells MD-MB-435.

METHODS

MD-MB-435 cells were sense-transfected with ST6Gal I cDNA or antisense-transfected with part of the ST6Gal I sequence inserted in pcDNA 3.1 vector, with mock transfection with pcDNA3.1 vector as the control. The cell surface alpha2, 6-linked sialylation was determined by fluorescence-activated cell sorting (FACS) using lectin SNA (Sambucus nigra agglutinin specific to alpha2, 6-linked sialic acid on N-linked glycoprotein). A significantly increased alpha2, 6-sialylation subclone in sense-transfectants and a decreased alpha2, 6-sialylation subclone in antisense-transfectants were selected for further examination of cell-cell and cell-matrix (collagen IV) adhesion. The transfectants were also treated with sialidase to compare the capacity of cell adhesion affected by cell surface sialylation.

RESULTS

Sense-transfection subclone showed a reduced cell-cell aggregation but enhanced cell-matrix adhesion. In contrast, the antisense-transfection subclone exhibited increased cell-cell aggregation and decreased cell-matrix adhesion. After treatment with sialidase, the cell-matrix adhesion of all the transfectants and the parental MDA-MB-435 cells were significantly reduced to the level of 31%-57% of untreated cells.

CONCLUSION

Cell surface sialic acid and alpha2, 6-linked sialylation play an important role in cell-cell and cell-matrix adhesion of mammary carcinoma cell MDA-MB-435.

摘要

目的

研究细胞表面唾液酸及其连接方式对乳腺癌细胞MD-MB-435细胞间及细胞与基质黏附的影响。

方法

将MD-MB-435细胞用ST6Gal I cDNA进行正义转染,或用插入pcDNA 3.1载体的部分ST6Gal I序列进行反义转染,以pcDNA3.1载体的 mock转染作为对照。使用凝集素SNA(黑接骨木凝集素,对N-连接糖蛋白上的α2,6-连接唾液酸具有特异性)通过荧光激活细胞分选(FACS)测定细胞表面α2,6-连接唾液酸化。选择正义转染子中α2,6-唾液酸化显著增加的亚克隆和反义转染子中α2,6-唾液酸化降低的亚克隆,进一步检测细胞间及细胞与基质(IV型胶原)的黏附。转染子还用唾液酸酶处理,以比较细胞表面唾液酸化对细胞黏附能力的影响。

结果

正义转染亚克隆显示细胞间聚集减少,但细胞与基质的黏附增强。相反,反义转染亚克隆表现出细胞间聚集增加,细胞与基质的黏附减少。用唾液酸酶处理后,所有转染子及亲本MDA-MB-435细胞的细胞与基质黏附均显著降低至未处理细胞水平的31%-57%。

结论

细胞表面唾液酸及α2,6-连接唾液酸化在乳腺癌细胞MDA-MB-435的细胞间及细胞与基质黏附中起重要作用。

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