Regulation of Bub1 mRNA expression in endometria1 carcinoma Ishikawa cells by estrogen and Paclitaxel
10.3760/cma.j.issn.0529-567x.2010.09.011
- VernacularTitle:雌激素和紫杉醇对子宫内膜癌Ishikawa细胞中Bub1 mRNA表达的影响
- Author:
Yonghua CHEN
;
Xiaoping LI
;
Yue WANG
;
Jianliu WANG
;
Lihui WEI
- Publication Type:Journal Article
- Keywords:
Endometrial neoplasms;
Mitotic spindle apparatus;
Protein-serine-threonine kinases;
Estradiol;
Paclitaxel
- From:
Chinese Journal of Obstetrics and Gynecology
2010;45(9):686-690
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the regulation of Bub1 mRNA expression in endometrial carcinoma cells by estrogen and paclitaxel. Methods The high differentiated endometrial adenocarcinoma cells ( Ishikawa cell line) were cultured in DMED/F12 supplemented with 10% fetal bovine serum(FBS) or phenol red-free DMED/F12 supplemented with 5% dextran-charcoal FBS (dcFBS). Firstly, the cells were stimulated by 10 nmol/L estradiol (17β-E2 ) or no hormonal stimulation as control group, and the cell proliferation was quantified at 24, 48 and 72 hours using cell counting kit-8 (CCK-8) method. Then the cells were stimulated with different concentrations of 17β-E2 (0.1, 10, 1000 nmol/L) at different periods (5,15,30 minites and 2,4,8,12,16,24,30 hours), the expression of Bub1 mRNA was detected by real-time quantitative PCR. Ishikawa cells were cultured with non-serum DMEM/F12 to be synchronized, and then were treated with different concentrations of paclitaxel( 10,100 nmol/L) for 8 and 24 hours. While, nonsynchronized Ishikawa cells were exposed to 100 nmoL/L paclitaxel for different periods(4, 8, 16, 24,48 hours), and real-time quantitative PCR was also used to detect the expression levels of Bub1 mRNA.Data were presented as folds change relative to control group, in which values ＜ 1 were down-regulated, and those ＞ 1 were up regulated. Results The proliferation rate of cells in the presence of 17β-E2 was significantly highter than that of the control group after treated 24 hours (A value: 0. 70 ±0. 08 vs. 0. 86 ±0.10, P = 0.049). Time-dependent experiments revealed that addition of 17β-E2 could increase cell numbers during 72 hours period, while the expression level of Bub1 mRNA was decreased in Ishikawa cell.Dose-dependent experiments revealed maximal estradiol stimulation effects at 10 nmol/L( P = 0. 020). After being treated with serum-free culture, Ishikawa cells were exposed to 10 nmol/L paclitaxel for 8 and 24 hours, and the expression of Bubl mRNA decreased (0. 403 ± 0. 008 vs. 0. 775 ± 0. 144, P = 0. 251 ).Compared to the control cells, the mRNA expression levels of Bub1 in cells treated by paclitaxel for 8 hours was significantly decreased ( P = 0. 009 ), while there was not significantly decreased at 24 hours ( P =0. 396). When exposed to 100 nmol/L paclitaxel for 8 and 24 hours, the expression of Bubl mRNA was also decreased(0. 697 ±0. 017 vs. 0. 850 ±0. 004, P =0. 061 ). Compared to the control cells, Bub1 mRNA expression was also significantly decreased (P = 0. 038 and P= 0. 019, respectively). While with serum freetreatment culture, when Ishikawa cells exposed to 100 nmoL/L paclitxel for 4, 8, 16, 24 or 48 hours, the expression of Bub1 mRNA significantly increased ( 1. 127 ± 0. 105 vs. 1. 614 ± 0. 154 vs. 2. 092 ± 0. 179vs. 1. 381 ± 0. 061 vs. 1. 519 ± 0. 182, P = 0. 002 ), of which was signicantaly increased at 16 hours treatment. Conclusion Bub1 exrpession could be regulated by estradiol and paclitaxel, in which deregulated Bubl expression may contribute to chemotherapeutic efficacy of paclitaxel.
