Xie Y L, Yang Y J, Tang C, Sheng H J, Jiang Y, Han K, Ding L J
Department of Obstetrics and Gynecology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Zhong Shan Road 321#, Nanjing, 210008, China,
Clin Transl Oncol. 2014 Oct;16(10):898-905. doi: 10.1007/s12094-014-1166-x. Epub 2014 Mar 19.
This study evaluated the effect of estrogen (E(2)), progesterone (P4), and the combination of them (E(2) + P4) on survival rate, apoptosis, and the expressions of Bcl-2, hsa-let-7a and has-miR-34b in primary ovarian cancer cells to provide new clues for the clinical treatments of ovarian cancer.
The primary ovarian cancer cells from 60 cases of clinical ovarian cancer tissues were isolated and then cultured. The survival rate of ovarian cancer cells after the treatment of E(2), P4 and E(2) + P4 was analyzed by MTT assay. Cell apoptosis rate and cell cycle were measured by FACS analysis. Moreover, the relative abundance of Bcl-2 and microRNAs (let-7a, miR-34b) expressions were detected by quantitative real-time PCR (qRT-PCR) and Western blotting.
Low concentrations of estrogen (10(-10), 10(-8), 10(-6 )mol/L) did not affect the proliferation of ovarian cancer cells. However, the high concentration of estrogen (10(-4 )mol/L) inhibited survival rate of ovarian cancer cells. Progesterone (10(-4 )mol/L) inhibited the proliferation of cancer cells. The combination of estrogen and progesterone significantly inhibited the survival rate of ovarian cancer cells with a time- and dose-dependent manner. High concentration of estrogen combined with progesterone (E(2) + P4) induced apoptosis of ovarian cancer cells. E(2) + P4 promoted the expression of let-7a and miR-34b and reduced the expression of Bcl-2 in ovarian cancer cells. When the expression of let-7a or/and miR-34b was inhibited using miRNA inhibitors, E(2) + P4 treatment did not change the protein level of Bcl-2.
E(2) + P4 significantly inhibited the cell survival, promoted the cell apoptosis, induced the expression of let-7a and miR-34b, and reduced the expression of Bcl-2 in ovarian cancer cells.
本研究评估雌激素（E₂）、孕激素（P4）及其联合应用（E₂ + P4）对原发性卵巢癌细胞存活率、凋亡以及Bcl-2、hsa-let-7a和has-miR-34b表达的影响，为卵巢癌的临床治疗提供新线索。
从60例临床卵巢癌组织中分离出原发性卵巢癌细胞并进行培养。采用MTT法分析E₂、P4和E₂ + P4处理后卵巢癌细胞的存活率。通过流式细胞术分析检测细胞凋亡率和细胞周期。此外，采用定量实时PCR（qRT-PCR）和蛋白质免疫印迹法检测Bcl-2和微小RNA（let-7a、miR-34b）表达的相对丰度。
低浓度雌激素（10⁻¹⁰、10⁻⁸、10⁻⁶ mol/L）不影响卵巢癌细胞的增殖。然而，高浓度雌激素（10⁻⁴ mol/L）抑制卵巢癌细胞的存活率。孕激素（10⁻⁴ mol/L）抑制癌细胞的增殖。雌激素和孕激素联合应用以时间和剂量依赖的方式显著抑制卵巢癌细胞的存活率。高浓度雌激素联合孕激素（E₂ + P4）诱导卵巢癌细胞凋亡。E₂ + P4促进卵巢癌细胞中let-7a和miR-34b的表达并降低Bcl-2的表达。当使用微小RNA抑制剂抑制let-7a或/和miR-34b的表达时，E₂ + P4处理并未改变Bcl-2的蛋白水平。
E₂ + P4显著抑制卵巢癌细胞的存活，促进细胞凋亡，诱导let-7a和miR-34b的表达，并降低Bcl-2在卵巢癌细胞中的表达。
