Low R B, Chaponnier C, Gabbiani G
Lab Invest. 1981 Apr;44(4):359-67.
To study the distribution and the degree of polymerization of actin in regenerating and neoplastic cells we have examined: (1) the immunofluorescent staining of these cells with human antiactin antibodies (AAA), (2) the sensitivity of cellular actin to AAA staining after treatment with the actin-depolymerizing factor present in plasma or serum of several animal species, and (3) the total and relative amounts of F- and G-actin in tissue preparations, determined by means of differential centrifugation and densitometric analysis of stained sodium dodecyl sulfate-polyacrylamide gels. There was little or no difference in staining intensity between normal and regenerating or tumoral tissues when treated with AAA, but AAA staining of normal tissues was abolished, whereas staining of regenerating and tumoral tissues remained intense after incubation with actin-depolymerizing factor. Gel analysis showed that normal, regenerating, and neoplastic epidermal cells contained similar amounts of actin. Compared to controls, regenerating liver tissue contained slightly more total actin, nearly the same amount of G-actin, but a substantially increased amount of F-actin. These results suggest that: (1) compared to controls, a greater proportion of regenerating and neoplastic tissue actin is stabilized against the action of actin-depolymerizing factor; (2) changes involving actin in regenerating and neoplastic epithelial cells reflect changes in its degree of polymerization rather than its total amount.
为了研究肌动蛋白在再生细胞和肿瘤细胞中的分布及聚合程度,我们进行了以下检测:(1)用人类抗肌动蛋白抗体(AAA)对这些细胞进行免疫荧光染色;(2)用几种动物血浆或血清中存在的肌动蛋白解聚因子处理细胞后,检测细胞肌动蛋白对AAA染色的敏感性;(3)通过差速离心和对十二烷基硫酸钠-聚丙烯酰胺凝胶染色后的光密度分析,测定组织制剂中F-肌动蛋白和G-肌动蛋白的总量及相对含量。用AAA处理时,正常组织与再生组织或肿瘤组织之间的染色强度几乎没有差异,但正常组织的AAA染色被消除,而再生组织和肿瘤组织在用肌动蛋白解聚因子孵育后染色仍很强烈。凝胶分析表明,正常、再生和肿瘤表皮细胞含有的肌动蛋白量相似。与对照组相比,再生肝组织的总肌动蛋白量略有增加,G-肌动蛋白量几乎相同,但F-肌动蛋白量大幅增加。这些结果表明:(1)与对照组相比,再生组织和肿瘤组织中更大比例的肌动蛋白对肌动蛋白解聚因子的作用具有稳定性;(2)再生和肿瘤上皮细胞中涉及肌动蛋白的变化反映了其聚合程度的变化,而非总量的变化。