主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

JOURNAL OF BEIJIGN UNIVERSITY OF TRADITIONAL CHINE ›› 2018, Vol. 41 ›› Issue (11): 935-942.doi: 10.3969/j.issn.1006-2157.2018.11.009

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Influence of genistein in high and low concentration on growth and ERα/ERβ protein ratio of Ishikawa cells*

Cheng Ran, Liu Xiaoli, Xue Xiao’ou#, Xie Wei   

  1. Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
  • Received:2018-07-16 Online:2018-11-30 Published:2018-11-30
  • Supported by:
    National Natural Science Foundation of China (No.81603430)

Abstract: Objective To study the difference of genistein between high and low concentration on the proliferation, colony formation and protein expression ratio of ERα to ERβ (ERα/ERβ) in Ishikawa cells of human endometrial carcinoma with positive estrogen receptor (ER). Methods ER-positive Ishikawa cells of human endometrial carcinoma were taken as medium research objects. Before formal experiments, the cells were cultured in Opti-MEM phenol-free serummedium for 24 h, and then divided into low-dose genistein group, high-dose genistein group and solvent control group. The genistein solution and solvent control were prepared by the Opti-MEM phenol-free serum medium containing 2% DCS-FBS and were sequentially added to 3 groups. The influence of high-concentration genistein (10 μmoL/L, 25 μmoL/L, 50 μmoL/L) and low-concentration genistein (0.001 μmoL/L, 0.01 μmoL/L, 0.1 μmoL/L, 1 μmoL/L) on proliferation of Ishikawa cells was detected by using cell counting kit-8 (CCK-8) after intervening for 24 h, 48 h and 72 h. The influence of high-concentration genistein (25 μmoL/L, 50 μmoL/L) and low-concentration genistein (0.01 μmoL/L, 0.1 μmoL/L) on colony formation was detected by using colony formation assay after intervening for 1 week. The influence of high-concentration genistein (25 μmoL/L, 50 μmoL/L) and low-concentration genistein (0.01 μmoL/L, 0.1 μmoL/L) on protein expression ratio of ERα/ERβ by using Western blotting assay after intervening for 48 h. Results Low-concentration genistein (0.001 μmoL/L, 0.01 μmoL/L, 0.1 μmoL/L) promoted the proliferation of Ishikawa cells, which was the most significant after intervening for 48 h, and proliferation promoting effect was the highest when genistein concentration reached 0.1 μmoL/L (P<0.01). High-concentration genistein 10 μmoL/L, 25 μmoL/L, 50 μmoL/L inhibited the proliferation of Ishikawa cells and showed time-concentration dependence. The inhibitory effect reached the highest when genistein concentration was 50 μmoL/L after 72 h. The colony formation rate was 14.60% and 15.13% respectively in low-concentration genistein groups compared with solvent control group (12.20%, P<0.05). The colony formation rate was 6.07% and 4.27% respectively in high-concentration genistein groups, which was significantly lower than that in solvent control group (P<0.01). The protein expression of ERα was up-regulated in high-concentration genistein groups and low-concentration genistein groups, and the expression reached the highest when genistein concentration was 0.1 μmoL/L. The protein expression of ERβ was down-regulated, and the expression reached the lowest when genistein concentration was 25 μmoL/L or 50 μmoL/L. The protein expression ratio of ERα/ERβ was significantly up-regulated, and reached the highest when genistein concentration was 0.1 μmoL/L. Conclusion Low-concentration genistein has effect of promoting Ishikawa cells with positive ER, which may be related to up-regulation of protein expression ratio of ERα/ERβ. High-concentration genistein can also up-regulate protein expression ratio of ERα/ERβ, but may play the role of inhibiting growth of Ishikawa cells through non-ER dependent mechanism.

Key words: phytoestrogen, genistein, Ishikawa cell line, endometrial carcinoma, ratio of estrogen receptor α to estrogen receptor β, proliferation, colony formation

CLC Number: 

  • R285.5