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转化生长因子β诱导小鼠乳头瘤细胞中寻常型天疱疮抗原活性

Induction by transforming growth factor beta of pemphigus vulgaris antigen activity in mouse papilloma cells.

作者信息

Kulesz-Martin M, Glurich I, Lisafeld B, Kozlowski P

机构信息

Grace Cancer Drug Center, Roswell Park Memorial Institute, Buffalo, New York 14263.

出版信息

Cancer Res. 1990 Feb 1;50(3):686-90.

PMID:2404575
Abstract

Induction of a marker of epidermal spinous cells, pemphigus antigen activity, was detected by indirect immunofluorescence in murine papilloma cells exposed to human transforming growth factor beta (TGF-beta). Detection of pemphigus antigen activity required exposure of cells to 1.4 mM Ca2+ for 3 h just prior to immunoassay. The brief exposure to Ca2+ may be necessary for translocation of intracellular pemphigus antigen to the cell surface, where it is accessible to antibody. Cells grown in medium containing 0.02-0.04 mM Ca2+ were shown previously to be primarily basal cells characterized by pemphigoid antigen activity. Following treatment with 0.25-25 pg/ml TGF-beta for 44 h under 0.02-0.04 mM Ca2+ conditions, 63 +/- 9% (SD) of cells were pemphigus positive. This percentage was comparable to that of positive control cultures exposed to 1.4 mM Ca2+ for 44 h (70 +/- 10%) and was up to 2-fold that of solvent control cultures. Pemphigus antigen activity was significantly induced by 0.1-25 pg/ml TGF-beta, out of a tested range of 10(-5)-10(3) pg/ml. The total number of papilloma cell colonies was unaffected by treatment with 0.1-25 pg/ml TGF-beta but was reduced greater than 90% by treatment with 10(3)-5 x 10(3) pg/ml TGF-beta. The described immunofluorescence assay for pemphigus antigen activity may be useful for preliminary evaluation of differentiation-inducing agents in anticarcinoma therapy.

摘要

在暴露于人类转化生长因子β(TGF-β)的小鼠乳头瘤细胞中,通过间接免疫荧光检测到了表皮棘细胞标志物天疱疮抗原活性的诱导。在免疫测定前,细胞需要暴露于1.4 mM Ca2+ 3小时才能检测到天疱疮抗原活性。短暂暴露于Ca2+可能是细胞内天疱疮抗原转运到细胞表面所必需的,在细胞表面抗体可以与之结合。先前已证明,在含有0.02 - 0.04 mM Ca2+的培养基中生长的细胞主要是具有类天疱疮抗原活性的基底细胞。在0.02 - 0.04 mM Ca2+条件下,用0.25 - 25 pg/ml TGF-β处理44小时后,63±9%(标准差)的细胞天疱疮呈阳性。该百分比与在1.4 mM Ca2+下暴露44小时的阳性对照培养物(70±10%)相当,是溶剂对照培养物的2倍。在10(-5)-10(3) pg/ml的测试范围内,0.1 - 25 pg/ml TGF-β可显著诱导天疱疮抗原活性。0.1 - 25 pg/ml TGF-β处理对乳头瘤细胞集落总数无影响,但10(3)-5×10(3) pg/ml TGF-β处理可使细胞集落总数减少90%以上。所描述的天疱疮抗原活性免疫荧光测定法可能有助于抗癌治疗中诱导分化剂的初步评估。

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