| 摘要: |
| 目的 探讨盐屋霉素A下调叉头框蛋白M1（forkhead box protein M1，FoxM1）对喉癌细胞增殖、凋亡、侵袭能力等恶性生物学行为的影响。方法 取处于对数生长期的Hep-2细胞，使用不同浓度盐屋霉素A进行处理，对照组不加盐屋霉素A。分别在培养24、48、72 h后通过CCK-8法检测细胞活力；24、48 h后通过CFSE染色法检测细胞增殖能力；48 h后Annexin Ⅴ-FITC/PI双染法检测细胞凋亡情况，Transwell小室法检测细胞侵袭能力。蛋白质印迹法检测FoxM1、Ki-67、cleaved caspase-3蛋白的表达变化，ELISA法检测细胞上清液中基质金属蛋白酶（MMP）-2、MMP-9的表达变化。结果 （1）盐屋霉素A作用后FoxM1蛋白的表达量降低（P<0.05）。CCK-8检测结果显示不同浓度盐屋霉素A对Hep-2细胞活力均有抑制作用，并随着盐屋霉素A浓度的增加和作用时间的延长，Hep-2的细胞活力逐渐下降，呈现浓度和时间依赖性。（2）1.3、1.5 μmol/L盐屋霉素A处理后Hep-2细胞的增殖能力均降低（P<0.05），并呈现浓度依赖性；同时Ki-67蛋白的表达下调。（3）1.3、1.5 μmol/L盐屋霉素A作用后，Hep-2细胞的凋亡率（10.6%，27.9 %）均高于对照组（4.91%，P<0.05），并呈药物浓度依赖性；同时 cleaved caspase-3蛋白的表达上调。（4）1.3、1.5 μmol/L盐屋霉素A作用后Hep-2细胞的侵袭能力较对照组均下降（P<0.05），并呈药物浓度依赖性；同时 MMP-2、MMP-9蛋白的表达下调。结论 盐屋霉素A下调FoxM1可抑制喉癌细胞的增殖、侵袭等生物学行为。 |
| 关键词: 喉肿瘤 盐屋霉素A 叉头框蛋白M1 细胞增殖 肿瘤侵袭 |
| DOI：10.16781/j.0258-879x.2016.08.0963 |
| 投稿时间：2015-12-16修订日期：2016-06-12 |
| 基金项目:国家自然科学基金面上项目（81272980），国家临床重点专科建设项目[卫办医政函（2012）649号]. |
|
| Down-regulation of forkhead box protein M1 by siomycin A can inhibit the malignant behaviors of laryngeal carcinoma cells |
| JIANG Li-zhu,LIU Ya-nan,WEN Tao-yu,CHEN Hong-yan* |
(Department of Otolaryngology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the effect of siomycin A-induced forkhead box protein M1(FoxM1) down-regulation on the malignant behaviors (proliferation, apoptosis, and invasive ability) of laryngeal carcinoma cells. Methods The experiment was performed with Hep-2 cells of the logarithmic growth phase. The experimental groups were treated with different doses of siomycin A, and the control group was treated without siomycin A. CCK-8 test was used to detect the viability of Hep-2 cells after treatment for 24, 48, and 72 h. CFSE staining was used to examine the proliferation ability of Hep-2 cells after treatment for 24 and 48 h. Annexin Ⅴ-FITC/PI dual staining was used to observe the siomycin A-induced apoptosis and Transwell test was used to examine the invasion ability of Hep-2 cells after treatment for 48 h. The expressions of FoxM1, Ki-67 and cleaved caspase-3 protein in Hep-2 cells were detected by Western blotting analysis. The supernatant levels of matrix metalloproteinase (MMP)-2 and MMP-9 were examined by ELISA assay. Results (1) Treatment with siomycin A significantly decreased the expression of FoxM1 protein compared with the control group (P<0.05). CCK-8 test found that siomycin A of different concentrations suppressed the viability of Hep-2 cells in a time- and dose-dependent manner. (2) Siomycin A at 1.3 and 1.5 μmol/L significantly suppressed the proliferation of Hep-2 cells in a dose-dependent manner (P<0.05), accompanied by down-regulated expression of Ki-67.(3)Siomycin A at 1.3 and 1.5 μmol/L also induced greater apoptosis of Hep-2 cells (10.6%, 27.9 %) in a dose-dependent manner compared with control group (4.91%, P<0.05), accompanied by up-regulated cleaved caspase-3 expression. (4) Siomycin A at 1.3 and 1.5 μmol/L suppressed the invasive ability of Hep-2 cells in a dose-dependent manner compared with control group (P<0.05), accompanied by down-regulated expression of MMP-2 and MMP-9. Conclusion Down-regulation of FoxM1 by siomycin A can inhibit the proliferation and invasive ability of laryngeal carcinoma cells. |
| Key words: laryngeal neoplasmas siomycin A forkhead box protein M1 cell proliferation neoplasm invasiveness |
