Adler R A, Naumann S A, Mansouri A, Krieg R J, Latta K, Sanders K M
Section of Endocrinology, McGuire Veterans Affairs Medical Center, Richmond, VA 23249.
Life Sci. 1994;55(23):1823-31. doi: 10.1016/0024-3205(94)90093-0.
Prolactin and other lactogenic hormones are mitogenic for the rat T-cell lymphoma line, Nb2. Glucocorticoids have antiproliferative effects on these cells. A limiting feature of experiments utilizing the Nb2 line is their labor-intensive nature. We therefore adapted the commonly used MTT dye proliferation assay for the Nb2 cell line. While rPRL, hPRL, oPRL, hGH, bPL, and to a lesser extent bPRL stimulated the Nb2 cells, hormones without lactogenic activity, rGH and oGH did not. Human serum and rat sera from animals bearing a PRL-secreting tumor stimulated the Nb2 cells in parallel to standards. Glucocorticoids had anti-proliferative effects on Nb2 cells in the presence of half-maximal or maximal PRL doses, as measured by the MTT proliferation assay. It has been claimed that an oxazoline steroid, deflazacort, has anti-inflammatory effects in clinical studies with fewer of the deleterious side-effects common to glucocorticoids. We therefore compared the in vitro anti-proliferative effects of deflazacort with other glucocorticoids. Deflazacort's negative effect on Nb2 cell proliferation was similar to that of cortisol and prednisolone and less than that of dexamethasone. We conclude that the MTT proliferation assay can be used to study both mitogenic and anti-proliferative substances in Nb2 cells. In addition we found that deflazacort acts similarly in vitro to other glucocorticoids.
催乳素及其他生乳激素对大鼠T细胞淋巴瘤细胞系Nb2具有促有丝分裂作用。糖皮质激素对这些细胞具有抗增殖作用。利用Nb2细胞系进行实验的一个局限性在于其实验过程较为繁琐。因此,我们对常用的MTT染料增殖试验进行了调整,使其适用于Nb2细胞系。虽然重组人催乳素(rPRL)、人催乳素(hPRL)、羊催乳素(oPRL)、人生长激素(hGH)、牛催乳素(bPL),以及在较小程度上的牛催乳素(bPRL)能刺激Nb2细胞,但无生乳活性的激素,即大鼠生长激素(rGH)和羊生长激素(oGH)则不能。来自患有分泌催乳素肿瘤动物的人血清和大鼠血清与标准品一样能刺激Nb2细胞。通过MTT增殖试验测定,在半最大或最大催乳素剂量存在的情况下,糖皮质激素对Nb2细胞具有抗增殖作用。据称,一种恶唑啉类甾体药物,即地夫可特,在临床研究中具有抗炎作用,且糖皮质激素常见的有害副作用较少。因此,我们比较了地夫可特与其他糖皮质激素在体外的抗增殖作用。地夫可特对Nb2细胞增殖的负面影响与皮质醇和泼尼松龙相似,且小于地塞米松。我们得出结论,MTT增殖试验可用于研究Nb2细胞中的促有丝分裂和抗增殖物质。此外,我们发现地夫可特在体外的作用与其他糖皮质激素相似。