Yoon H S, Rho S H, Jeong J H, Yoon S, Yoo K S, Yoo Y H
Department of Ophthalmology, Institute of Medical Science, Dong-A University College of Medicine, Pusan, South Korea.
Curr Eye Res. 2000 Mar;20(3):215-24.
To investigate the effect of the tyrosine kinase inhibitor, genistein, on the growth of the retinal pigment epithelial (RPE) cell.
The tyrosine kinase inhibitor, genistein, was administered in culture to the rat retinal pigment epithelial cell line, RPE-J. The effect on cell viability and growth was assessed by trypan blue dye exclusion. Induction of apoptosis was confirmed morphologically by light and electron microscopy and oligonucleosomal fragmentation was assessed by TUNEL and DNA ladder. Quantitation was undertaken by propidium iodide staining and photometric enzyme immunoassay. Western blot was performed to study poly-(ADP-ribose)-polymerase cleavage (PARP). To confirm the involvement of caspase, the caspase inhibitor z-VAD-fmk was employed. In addition, cell cycle phase was determined by flow cytometry.
We here demonstrate that genistein treatment of RPE-J cells produces a dose- and time-dependent growth inhibition. Genistein in higher concentration induces apoptosis of rat RPE-J cell. z-VAD-fmk inhibited this type of apoptosis and cleavage of PARP enzyme was demonstrated. Ten micromolar genistein inhibited cell proliferation by G(0)/G(1) arrest without inducing apoptosis of the major population. Whereas 50 microM genistein caused growth inhibition of RPE-J cells by G(2)/M arrest and subsequent apoptotic death.
Genistein inhibits RPE cell growth and induces apoptosis. The ability of genistein to inhibit the proliferation and to induce apoptosis of RPE cells could be potentially therapeutic for proliferative vitreoretinopathy.
研究酪氨酸激酶抑制剂染料木黄酮对视网膜色素上皮（RPE）细胞生长的影响。
将酪氨酸激酶抑制剂染料木黄酮应用于培养的大鼠视网膜色素上皮细胞系RPE-J。通过台盼蓝拒染法评估其对细胞活力和生长的影响。通过光镜和电镜从形态学上确认凋亡的诱导情况，并通过TUNEL和DNA梯状条带评估寡核小体片段化。通过碘化丙啶染色和光度酶免疫测定进行定量分析。进行蛋白质印迹法以研究聚（ADP-核糖）聚合酶裂解（PARP）。为了确认半胱天冬酶的参与，使用了半胱天冬酶抑制剂z-VAD-fmk。此外，通过流式细胞术确定细胞周期阶段。
我们在此证明，染料木黄酮处理RPE-J细胞会产生剂量和时间依赖性的生长抑制。较高浓度的染料木黄酮可诱导大鼠RPE-J细胞凋亡。z-VAD-fmk抑制了这种类型的凋亡，并证明了PARP酶的裂解。10微摩尔的染料木黄酮通过G(0)/G(1)期阻滞抑制细胞增殖，而不诱导主要群体的凋亡。而50微摩尔的染料木黄酮通过G(2)/M期阻滞和随后的凋亡死亡导致RPE-J细胞生长抑制。
染料木黄酮抑制RPE细胞生长并诱导凋亡。染料木黄酮抑制RPE细胞增殖和诱导凋亡的能力可能对增殖性玻璃体视网膜病变具有潜在的治疗作用。
