Paik K J, Jeon S S, Chung H Y, Lee K H, Kim K W, Chung J K, Kim N D
Department of Pharmacy, Pusan National University, Korea.
Arch Pharm Res. 1998 Aug;21(4):398-405. doi: 10.1007/BF02974633.
We investigated the effects of triterpene acids (TAs), ursolic acid (UA) and oleanolic acid (OA), on the induction of proliferation and differentiation of normal rat mammary epithelial cells (RMEC) or organoids cultured in Matrigel or primary culture system. To elucidate the effects, we tested their differentiation inducing activities with intercellular communication ability, cell cycle patterns, induction of apoptosis, and morphological differentiation in the three dimensional extracellular culture system. To study the changes of RMEC subpopulation in culture, the cultured cells were isolated, immunostained with peanut lectin (PNA) and anti-Thy-1.1 antibody and then analyzed with flow cytometry. Four different subpopulations, such as PNA and Thy-1.1 negative cells (B-), PNA positive cells (PNA+), Thy-1.1 positive cells (Thy-1.1+), PNA and Thy-1.1 positive cells (B+), were obtained and the size of each subpopulation was changed in culture with time in the presence of TAs. Intercellular communication was observed in culture for 7 days in TAs-treated cells, but not in culture for 4 days with scrape-loading dye transfer technique. G2/M phase cells and the number of apoptotic population were increased in TAs-treated groups in cell cycle analyses. S phase fractions were reduced and the change of G1 phase cells was not observed. The colonies with distinct multicellular structures, such as stellate, ductal, webbed, squamous, lobulo-ductal colonies, were observed in Matrigel culture and the frequencies of each colony were changed in the presence of TAs. These results suggest that UA and OA have differentiation inducing effects on rat mammary epithelial cells in primary or in Matrigel culture.
我们研究了三萜酸（TAs）、熊果酸（UA）和齐墩果酸（OA）对正常大鼠乳腺上皮细胞（RMEC）在基质胶或原代培养系统中增殖和分化诱导的影响。为了阐明这些影响，我们在三维细胞外培养系统中，通过细胞间通讯能力、细胞周期模式、凋亡诱导和形态分化来测试它们的分化诱导活性。为了研究培养过程中RMEC亚群的变化，将培养的细胞分离，用花生凝集素（PNA）和抗Thy-1.1抗体进行免疫染色，然后用流式细胞术分析。获得了四个不同的亚群，即PNA和Thy-1.1阴性细胞（B-）、PNA阳性细胞（PNA+）、Thy-1.1阳性细胞（Thy-1.1+）、PNA和Thy-1.1阳性细胞（B+），并且在TAs存在的情况下，每个亚群的大小在培养过程中随时间变化。用刮擦加载染料转移技术观察到，TAs处理的细胞在培养7天时存在细胞间通讯，但在培养4天时未观察到。细胞周期分析显示，TAs处理组的G2/M期细胞和凋亡群体数量增加。S期比例降低，未观察到G1期细胞的变化。在基质胶培养中观察到具有明显多细胞结构的集落，如星状、导管状、网状、鳞状、小叶导管状集落，并且在TAs存在的情况下，每个集落的频率发生了变化。这些结果表明，UA和OA对原代培养或基质胶培养中的大鼠乳腺上皮细胞具有分化诱导作用。
