Competence Effect of PDGF on Ki-67 Antigen and DNA Contents, and Its Inhibition by Trichostatin-A and a Butylydene Phthalide BP-421 in Primary Smooth Muscle Cells of Rat Aorta by Flow Cytometry

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We investigated with flow cytometry the platelet-derived growth factor (PDGF)-induced competence in primary cultured smooth muscle cells (SMC) of rat thoracic aorta. A cytogram was obtained by a double staining technique with fluorescein isothiocyanate-conjugated mouse monoclonal antibody against the proliferation-associated nucleus antigen Ki-67,and propidium iodide for total DNA content. (1) The nearly confluent SMC after 6d-culture with 5% fetal bovine serum (FBS) were further cultured under serum starvation for 2d. (2) SMC was cultured with 5% FBS for 4d, and then with 5% FBS+trichostatin-A (TS-A) (1 μg/ml) for 2d. The cytogram showed the broadening of Ki-67 antigen signal for G_0/G_1 phase by TS-A compared with that of SMC under serum starvation, suggesting the existence of early and late phases of G_1.(3) After serum starvation, the preculture with PDGF (100 ng/ml) for 3h which was followed by a further 15 h-culture with 3% FBS caused significant more entry into S phase than control culture. The extent was greater than that with 15 h-culture with 10% FBS. (4) 15 h-culture with 1% FBS after PDGF (30 ng/ml) pretreatment stimulated entry into S phase cells, which was inhibited by TS-A (1 μg/ml) and by a butylydene phthalide derivative BP-421 (3 μg/ml). Flow cytometric analysis demonstrate that PDGF pretreatment stimulates the entry into S phase by 20% of total SMC having early and late phases of G_1,and that the PDGF-competence is inhibited by TS-A and BP-421.
公益社団法人日本薬学会の論文
1995-12-15

Competence Effect of PDGF on Ki-67 Antigen and DNA Contents, and Its Inhibition by Trichostatin-A and a Butylydene Phthalide BP-421 in Primary Smooth Muscle Cells of Rat Aorta by Flow Cytometry

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