棉酚对人子宫肌瘤和子宫平滑肌细胞生长抑制和凋亡的作用与 Bcl-2、Src 和 ERα 有关。
Involvement of Bcl-2, Src, and ERα in gossypol-mediated growth inhibition and apoptosis in human uterine leiomyoma and myometrial cells.
机构信息
Department of Reproductive Pharmacology, NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China.
出版信息
Acta Pharmacol Sin. 2010 Dec;31(12):1593-603. doi: 10.1038/aps.2010.153. Epub 2010 Nov 22.
AIM
to investigate the effect of gossypol on the growth of cultured human uterine leiomyoma and myometrial cells, the level of Bcl-2 and the activity of Src and estrogen receptor (ERα).
METHODS
human uterine leiomyoma and adjacent normal myometrial cells were cultured in vitro. Both cell types were treated with a graded concentration of gossypol. Cell viability was assayed using CCK-8. Morphological change was observed with optical and electronic microscopy. Apoptosis was evaluated using TUNEL assay. Levels of Bcl-2, ERα and Src were analyzed using Western blotting.
RESULTS
gossypol significantly inhibited growth and promoted apoptosis in cultured human uterine leiomyoma cells with the IC(50) value and its corresponding 95% confidence intervals (CI) of 6.5 (4.0-10.5), 9.0 (4.9-16.5), and 7.5 (4.0-14.1) micromol/L at 20, 40, and 60 h, respectively. Gossypol exerted inhibitory effects on the myometrial cells with the IC(50) value and its 95% CI of 49.1 (28.3-85.0), 14.5 (7.7-27.4), and 2.6 (1.2-5.6) micromol/L at 20, 40, and 60 h, respectively. Compared with control, gossypol 0.1-3.0 micromol/L markedly decreased the protein expression of Bcl-2 (P<0.05) in both leiomyoma and myometrial cells in a concentration-dependent manner, and significantly suppressed the level of phospho-Tyr416Src (P<0.05) in both cell types at 3.0 micromol/L without obvious alteration of c-Src and phospho-Tyr527Src levels (P>0.05). In addition, gossypol markedly reduced both the expression of ERα (P<0.05) at the low concentration of 0.1 micromol/L in the myometrial cells and the level of phospho-ser167ERα (P<0.05) at the high concentration of 3.0 μmol/L in the leiomyoma cells.
CONCLUSION
gossypol inhibits proliferation and induces apoptosis in human uterine leiomyoma and myometrial cells. It is likely that the mechanisms of action involve reducing the protein level of Bcl-2 and the activity of Src and ERα.
目的
探讨棉酚对体外培养的人子宫肌瘤及子宫平滑肌细胞生长、Bcl-2 水平及原癌基因 Src 和雌激素受体（ERα）活性的影响。
方法
体外培养人子宫肌瘤及邻近正常子宫平滑肌细胞，用不同浓度棉酚处理细胞。采用 CCK-8 法检测细胞活力，光镜和电镜观察细胞形态学变化，TUNEL 法检测细胞凋亡，Western blot 法检测 Bcl-2、ERα 和 Src 蛋白表达。
结果
棉酚显著抑制人子宫肌瘤细胞生长，促进其凋亡，作用 20、40、60 h 的 IC50 及其 95%可信区间（CI）分别为 6.5（4.0～10.5）、9.0（4.9～16.5）、7.5（4.0～14.1）μmol/L；对子宫平滑肌细胞也有抑制作用，作用 20、40、60 h 的 IC50 及其 95%CI 分别为 49.1（28.3～85.0）、14.5（7.7～27.4）、2.6（1.2～5.6）μmol/L。与对照组相比，棉酚 0.1～3.0 μmol/L 时，两种细胞中 Bcl-2 蛋白表达均明显降低（P＜0.05），且呈浓度依赖性；两种细胞中磷酸化 Tyr416Src（p-Tyr416Src）水平均明显降低（P＜0.05），但对 c-Src 和磷酸化 Tyr527Src（p-Tyr527Src）水平无明显影响（P＞0.05）。此外，棉酚 0.1 μmol/L 时可明显降低子宫平滑肌细胞中 ERα 表达（P＜0.05），3.0 μmol/L 时可明显降低子宫肌瘤细胞中磷酸化 Ser167 ERα（p-Ser167 ERα）水平（P＜0.05）。
结论
棉酚抑制人子宫肌瘤及子宫平滑肌细胞的增殖并诱导其凋亡，其作用机制可能与降低 Bcl-2 蛋白水平和 Src、ERα 活性有关。
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