Elsevier

Urology

Volume 64, Issue 2, August 2004, Pages 389-393
Urology

Basic science
Antiproliferative and antiangiogenic activities of genistein in human renal cell carcinoma

https://doi.org/10.1016/j.urology.2004.03.045Get rights and content

Abstract

Objectives

To determine whether genistein, an isoflavone that is plentiful in soy beans, could inhibit the growth of human renal cell carcinoma (RCC) cells in vitro, induce apoptosis, and suppress neovascularization in vivo induced by human RCC cells.

Methods

We investigated the effect of genistein on cell proliferation in four human RCC cell lines, SMKT R-1, R-2, R-3, and R-4. The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling assay was performed to examine whether genistein could induce apoptosis in SMKT R-1 cells. To evaluate the effect of genistein on in vivo angiogenesis, we used a new mouse dorsal air sac model in which we could evaluate it simply and quantitatively. Radioisotope-labeled red blood cells were injected into a tail vein in mice bearing a Millipore filter chamber containing genistein, and the vascular volume was examined by measuring the radioactivity of the mouse dorsal skin.

Results

Treatment with genistein for 48 hours inhibited cell proliferation in a dose-dependent manner and 100 μg/mL genistein inhibited it in a time-dependent manner. A dose of 50 μg/mL genistein clearly induced cell apoptosis. The vascular volume after implantation of the Millipore filter chamber containing RCC cells increased to threefold that without RCC cells. Genistein in the Millipore filter chamber significantly decreased the neovascularization induced by human RCC cells in vivo.

Conclusions

The results of this study demonstrated that genistein inhibited cell proliferation, induced apoptosis, and suppressed in vivo angiogenesis in human RCC cells. Genistein may be a promising antitumorigenic and antiangiogenic agent for the treatment and prevention of RCC.

Section snippets

Cell lines, cultures, and reagents

Four human RCC cell lines SMKT R-1, R-2, R-3, and R-4, which were established in our laboratory, were maintained in minimal essential medium with d-valine modification medium containing 10% fetal bovine serum. Each cell line was derived from a primary lesion of an individual patient with RCC. All cell lines were histopathologically proven to be of conventional RCC origin.12, 13 Genistein was obtained from Waco Pure Chemical Industries (Osaka, Japan).

Cell proliferation experiment

These four human RCC cell lines (3 × 103)

Effect of genistein on cell proliferation in human RCC cell lines

When the four human RCC cell lines, SMKT R-1, R-2, R-3, and R-4 were treated with several concentrations of genistein for 48 hours, it inhibited the growth of all four in a dose-dependent manner (Fig. 1A). A dose of 100 μg/mL genistein inhibited the growth in a time-dependent fashion (Fig. 1B; P <0.0001, Spearman rank correlation test).

Effect of genistein on apoptosis in human RCC cell lines

To determine the induction of apoptosis by genistein, the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay was performed. The

Comment

The inhibitory effect of genistein on the proliferation of cells in various cancers such as leukemia, neuroblastoma, rhabdomyosarcoma, prostate cancer, and bladder cancer has been reported in several studies.1, 3 For human RCC, only one study, by Balbaa et al.17 has reported that genistein suppresses the proliferation of SMKT R-3 cells, which we established. In the current study, genistein inhibited the proliferation of all four human RCC cell types in vitro, and this inhibition was both dose

Acknowledgements

To Drs. Taro Senba, Toshiaki Wakabayashi, Yasuhiro Funahashi, and Kentaro Yoshimatsu, Tsukuba Research Laboratories, Eisai Co., Ltd. for technical support.

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    This work was supported in part by a Grant-in-Aid from the Japanese Ministry of Education, Science, Sports and Culture, and the Stiftelsen Japanese-Swedish Cooperative Research Foundation.

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