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学科主题: 临床医学
题名:
Nifedipine induced autophagy through Beclin1 and mTOR pathway in endometrial carcinoma cells
作者: Bao Xiao-xia1; Xie Bu-shan2; Li Qi1; Li Xiao-ping1; Wei Li-hui1; Wang Jian-liu1
关键词: nifedipine ; L-type ; autophagy ; endometrial carcinoma
刊名: CHINESE MEDICAL JOURNAL
发表日期: 2012-09-05
DOI: 10.3760/cma.j.issn.0366-6999.2012.17.028
卷: 125, 期:17, 页:3120-3126
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Medicine, General & Internal
研究领域[WOS]: General & Internal Medicine
关键词[WOS]: INOSITOL 1,4,5-TRISPHOSPHATE RECEPTOR ; CALCIUM ; CA2+ ; TARGET
英文摘要:

Background Endometrial carcinoma is one of the most common female tract genital malignant tumors. Nifedipine, an L-type calcium channel antagonist can inhibit cell proliferation of carcinomas. Recent studies indicated that a rise in the free cytosolic calcium ([Ca2+](c)) was a potent inducer of autophagy. Here, we investigated the relationship between nifedipine and autophagy in Hec-1A cells.

Methods Cells were cultured with nifedipine (10 mu mol/L) and harvested at different times for counting cell number. MTT assay was applied to evaluate the cell viability and transwell assay to reveal cell migration. Apoptotic cells were detected with annexin V/PI assay. Then cells were treated with 3-methyladenine (3-MA) (2.5 mmol/L) for 0, 5, 15, 30, 60, and 120 minutes and the expression of the L-type calcium channel alpha1D (Cav1.3) protein was detected. At last, cells were cultured and assigned to four groups with different treatment: untreated (control group), 10 mu mol/L nifedipine (N group), 2.5 mmol/L 3-MA (3-MA group), and 10 mu mol/L nifedipine plus 2.5 mmol/L 3-MA (N+3MA group). Autophagy was detected with GFP-LC3 modulation by fluorescent microscopy, and expression of the autophagy-associated proteins (LC3, Beclin1 and P70s6K) by Western blotting and monodansylcadaverine (MDC) labeled visualization.

Results Proliferation of Hec-1A cells was obviously suppressed by nifedipine compared with that of the untreated cells for 24, 48, and 96 hours (P=0.000 for each day). The suppression of migration ability of the nifedipine-treated cells (94.0 +/- 8.2) was significantly different from that of the untreated cells (160.00 +/- 9.50, P=0.021). The level of early period cell apoptosis induced by nifedipine was (2.21 +/- 0.19)%, which was (2.90 +/- 0.13)% in control group (P=0.052), whereas the late period apoptosis level reached (10.38 +/- 0.96)% and (4.40 +/- 0.60)% (P=0.020), respectively. The 3-MA group induced a slight increase in the Cav1.3 levels within 15 minutes, but significantly attenuated the Cav1.3 levels after 30 minutes. There were more autophagic vacuoles labeled by MDC in the N group (20.63 +/- 3.36) than the control group (6.29 +/- 0.16, P=0.015). GFP-LC3 localization revealed that the LC3 levels of cells in 3-MA group, N+3MA group, 3-MA group were 2.80 +/- 0.29, 2.30 +/- 0.17, and 1.80 +/- 0.21, respectively. Cells in the N group showed significant augmentation of autophagy (P <0.05). Western blotting analysis confirmed the down-regulation of LC3 levels in 3-MA group (0.85 +/- 0.21) and N+3MA group (1.21 +/- 0.12) compared with nifedipine treatment (2.64 +/- 0.15, P <0.05). The annexin-V-FITC/PI assay showed that the level of early period cell apoptosis induced in the N+3-MA group ((11.22 +/- 0.91)%) differed significantly from that of the control group ((2.51 +/- 0.70)%) and N group ((3.47 +/- 0.39)%). Similarly, the late period level of the N+3-MA group ((55.19 +/- 2.51)%) differed significantly from that of the control group((15.81 +/- 1.36)%) and the N group ((22.09 +/- 2.48)%, P <0.05). The down-regulated expression of P70s6k and up-regulated expression of the Beclin1 revealed significant differences between the N+3-MA group and control group (P=0.025; Beclin1: P=0.015).

Conclusions Proliferation and migration in vitro of endometrial carcinoma Hec-1A cells are significantly suppressed by nifedipine. The nifedipine leads autophagy to oppose Hec-1A cells apoptosis. Autophagy inhibition by 3-MA leads down-regulation of Cav1.3 and enhances nifedipine-induced cell death. The nifedipine-induced autophagy is linked to Beclin1 and mTOR pathways. Chin Med J 2012;125(17):3120-3126

语种: 英语
所属项目编号: 30973182
项目资助者: National Natural Science Foundation of China
WOS记录号: WOS:000309378100029
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/63899
Appears in Collections:北京大学第二临床医学院_期刊论文

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作者单位: 1.Peking Univ, Peoples Hosp, Dept Obstet & Gynecol, Beijing 100044, Peoples R China
2.Nanchang Univ, Affiliated Hosp 1, Dept Gastroenterol, Nanchang 330006, Jiangxi, Peoples R China

Recommended Citation:
Bao Xiao-xia,Xie Bu-shan,Li Qi,et al. Nifedipine induced autophagy through Beclin1 and mTOR pathway in endometrial carcinoma cells[J]. CHINESE MEDICAL JOURNAL,2012,125(17):3120-3126.
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