三氧化二砷对子宫内膜癌醋酸甲羟孕酮耐药细胞的作用研究
发布时间：2019-06-19 12:54
【摘要】：目的研究三氧化二砷(AS_2O_3)对子宫内膜癌细胞和耐药细胞的作用及机制。方法 (1)分别采用MTS法和Annexin V-FITC/PI双染色流式细胞术检测不同浓度的AS_2O_3在不同作用时间下对ISK细胞增殖和凋亡的影响;(2)采用浓度梯度递增持续刺激诱导法,体外建立MPA耐药细胞后,采用MTS法和Annexin V-FITC/PI双染色流式细胞术,检测AS_2O_3对ISK/MPA增殖和凋亡的影响,并与ISK组对比。(3)Western-blot法检测AS_2O_3作用后ISK与ISK/MPA细胞内p-AKT、p-ERK1/2及Caspase-3、Bcl-2和Bax蛋白的表达变化。(4)建立裸鼠皮下移植瘤模型,腹腔注射2 mg/kg的AS_2O_3,观察裸鼠瘤体的体积变化及毒副作用。结果 (1)AS_2O_3对ISK细胞有生长抑制作用,且呈时间和浓度依赖性;AS_2O_3作用后,ISK的凋亡率呈浓度依赖性升高,48 h细胞凋亡率大于24 h(P0.05);(2)成功建立人子宫内膜癌MPA耐药细胞系;AS_2O_3对ISK/MPA细胞具有生长抑制和促凋亡作用,且呈时间和浓度依赖性;AS_2O_3对ISK细胞组的促凋亡作用强于ISK/MPA细胞组(P0.05),但生长抑制作用比较,差异无统计学意义(P0.05);(3)AS_2O_3作用后,ISK及ISK/MPA细胞内p-AKT、p-ERK1/2和Caspase-3表达降低,Bcl-2表达降低而Bax表达升高(P0.05);(4)成功建立裸鼠皮下移植瘤模型,AS_2O_3作用后,裸鼠瘤体体积明显减小(P0.05)。结论 AS_2O_3对ISK/MPA细胞有增殖抑制和促凋亡作用,机制可能为AS_2O_3可导致Akt、ERK1/2磷酸化水平的降低,抑制P13K/AKT通路和MPAK/ERK通路的激活及在下调Bcl-2表达的同时,上调Bax蛋白的表达,继而调节凋亡相关蛋白通路分子Caspase-3的表达发挥增殖抑制和促凋亡作用。
[Abstract]:Objective to study the effect and mechanism of arsenic trioxide (AS_2O_3) on endometrial cancer cells and drug-resistant cells. Methods (1) the effects of different concentrations of AS_2O_3 on the proliferation and apoptosis of ISK cells were detected by MS assay and Annexin V-FITC/PI double staining flow cytometry. (2) MPA resistant cells were established in vitro by continuous stimulation with increasing concentration gradient. The effects of AS_2O_3 on the proliferation and apoptosis of ISK/MPA were detected by MS method and Annexin V-FITC/PI double staining flow cytometry, and compared with those in ISK group. (3) the effects of ISK and ISK/MPA cells treated with AS_2O_3 were detected by Western-blot assay. (3) ISK and ISK/MPA cells were treated with AS_2O_3. The expression of Bcl-2 and Bax protein was changed. (4) the subcutaneous tumor model of nude mice was established. 2 mg/kg AS_2O_3, was injected intraperitoneally to observe the volume changes and toxic and side effects of the tumors in nude mice. Results (1) AS_2O_3 could inhibit the growth of ISK cells in a time-and concentration-dependent manner, and the apoptosis rate of ISK increased in a concentration-dependent manner, and the apoptosis rate of MPA cells was more than 24 h at 48 h (P 0.05); (2). AS_2O_3 could inhibit the growth and promote apoptosis of ISK/MPA cells in a time-and concentration-dependent manner. The effect of AS_2O_3 on apoptosis in ISK cell group was stronger than that in ISK/MPA cell group (P 0.05), but there was no significant difference in growth inhibition effect between ISK and ISK/MPA cells treated with AS_2O_3 (P < 0.05). The expression of p 鈮，
本文编号：2502358
[Abstract]:Objective to study the effect and mechanism of arsenic trioxide (AS_2O_3) on endometrial cancer cells and drug-resistant cells. Methods (1) the effects of different concentrations of AS_2O_3 on the proliferation and apoptosis of ISK cells were detected by MS assay and Annexin V-FITC/PI double staining flow cytometry. (2) MPA resistant cells were established in vitro by continuous stimulation with increasing concentration gradient. The effects of AS_2O_3 on the proliferation and apoptosis of ISK/MPA were detected by MS method and Annexin V-FITC/PI double staining flow cytometry, and compared with those in ISK group. (3) the effects of ISK and ISK/MPA cells treated with AS_2O_3 were detected by Western-blot assay. (3) ISK and ISK/MPA cells were treated with AS_2O_3. The expression of Bcl-2 and Bax protein was changed. (4) the subcutaneous tumor model of nude mice was established. 2 mg/kg AS_2O_3, was injected intraperitoneally to observe the volume changes and toxic and side effects of the tumors in nude mice. Results (1) AS_2O_3 could inhibit the growth of ISK cells in a time-and concentration-dependent manner, and the apoptosis rate of ISK increased in a concentration-dependent manner, and the apoptosis rate of MPA cells was more than 24 h at 48 h (P 0.05); (2). AS_2O_3 could inhibit the growth and promote apoptosis of ISK/MPA cells in a time-and concentration-dependent manner. The effect of AS_2O_3 on apoptosis in ISK cell group was stronger than that in ISK/MPA cell group (P 0.05), but there was no significant difference in growth inhibition effect between ISK and ISK/MPA cells treated with AS_2O_3 (P < 0.05). The expression of p 鈮，
本文编号：2502358
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