国际肿瘤学杂志

国际肿瘤学杂志 ›› 2018, Vol. 45 ›› Issue (8): 453-459.doi: 10.3760/cma.j.issn.1673422X.2018.08.002

• 论著 • 上一篇    下一篇

蛇床子素对人鼻咽癌CNE2细胞增殖、侵袭、迁移能力的影响

陈炯玉,庄轶轩,庄珊珊,彭琳   

  1. 515041 汕头大学医学院附属肿瘤医院肿瘤研究实验室（陈炯玉、庄轶轩），检验科（庄珊珊、彭琳）
  • 出版日期:2018-08-08 发布日期:2018-11-01
  • 通讯作者: 彭琳，Email: st_penglin@126.com E-mail: st_penglin@126.com
  • 基金资助:

     广东省医学科研基金（B2018222、B2018049）；汕头市科技计划（汕府科〔2018〕37号）

Effects of osthole on the proliferation, invasion and migration of nasopharyngeal carcinoma cells CNE2

Chen Jiongyu, Zhuang Yixuan, Zhuang Shanshan, Peng Lin   

  1. Cancer Research Laboratory, Cancer Hospital of Shantou University Medical College, Shantou 515041, China
  • Online:2018-08-08 Published:2018-11-01
  • Contact: Peng Lin E-mail: st_penglin@126.com
  • Supported by:

    Medical Science Foundation of Guangdong Province of China (B2018222, B2018049); Science and Technology Program of Shantou City of China (Science and Technology Bureau of Shantou Government 〔2018〕37)

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摘要： 目的研究蛇床子素对人鼻咽癌CNE2细胞增殖、侵袭、迁移能力的影响，并探讨其调节CNE2细胞上皮间质转化（EMT）的分子机制。方法体外培养CNE2细胞，以0、20、40、80 μg/ml蛇床子素分别处理CNE2细胞24、48 h后，采用四甲基偶氮唑蓝（MTT）法、Transwell实验分别检测其对细胞增殖、侵袭和迁移能力的影响，以0 μg/ml蛇床子素处理组为对照组。反转录聚合酶链反应（RTPCR）和Western blotting方法分别检测EMT标志物上皮钙黏着蛋白（Ecadherin）、波形蛋白（vimentin）及Wnt/βcatenin信号通路组分βcatenin、细胞周期蛋白D1（cyclin D1）的转录和蛋白表达水平。结果作用24 h和48 h后，0、20、40、80 μg/ml蛇床子素处理组细胞增殖抑制率分别为0.00%±0.00%、7.45%±0.87%、14.12%±2.29%、27.26%±0.43%和0.00%±0.00%、13.44%±0.84%、29.03%±0.78%、57.49%±1.70%，差异均有统计学意义（F=174.33，P<0.001；F=1 041.40，P<0.001），进一步两两比较，差异均具有统计学意义（均P＜0.01）。采用不同浓度的蛇床子素（0、20、40、80 μg/ml）处理CNE2细胞48 h后，细胞迁移细胞数分别为52.13±4.49、29.00±4.49、18.50±1.93、13.75±2.77，差异有统计学意义（F=200.37，P＜0.001），进一步两两比较，差异均具有统计学意义（均P＜0.01）。采用不同浓度的蛇床子素（0、20、40、80 μg/ml）处理CNE2细胞48 h后，细胞侵袭数分别为46.63±2.87、24.13±2.87、16.75±5.29、11.00±1.77，差异有统计学意义（F=131.92，P＜0.001），进一步两两比较，差异均具有统计学意义（均P＜0.01）。采用不同浓度的蛇床子素（0、20、40、80 μg/ml）作用于人鼻咽癌CNE2细胞48 h后，Ecadherin mRNA（1.00±0.13、2.61±0.03、3.12±0.09、3.60±0.06；F=20.92，P<0.001）和Ecadherin蛋白表达（0.22±0.03、0.35±0.01、0.60±0.04、0.82±0.03；F=178.63，P＜0.001）差异有统计学意义，进一步两两比较，差异均有统计学意义（均P＜0.05）。同时，采用不同浓度蛇床子素（0、20、40、80 μg/ml）处理CNE2细胞48 h后，vimentin的mRNA表达（1.00±0.12、0.68±0.03、0.56±0.01、0.40±0.09；F=9.48，P<0.010）、βcatenin的mRNA表达（1.00±0.14、0.78±0.04、0.69±0.07、0.46±0.12；F=4.84，P<0.050）和cyclin D1的mRNA表达（1.00±0.09、0.82±0.03、0.58±0.09、0.40±0.03；F=9.49，P<0.010）差异均有统计学意义，各组间进一步两两比较，差异均有统计学意义（均P＜0.05）；vimentin的蛋白表达（0.85±0.02、0.74±0.01、0.34±0.01、0.27±0.01；F=610.58，P<0.001）、βcatenin的蛋白表达（0.83±0.00、0.44±0.02、0.39±0.00、0.23±0.03；F=985.74，P＜0.001）、cyclin D1的蛋白表达（0.86±0.02、0.67±0.00、0.35±0.01、0.25±0.01；F=910.57，P＜0.001）差异均有统计学意义，各组间进一步两两比较，差异均有统计学意义（均P＜0.05）。结论蛇床子素可抑制CNE2细胞的增殖、侵袭和迁移，其机制可能与调节Wnt/βcatenin信号通路进而抑制EMT过程有关。

关键词: 蛇床子素, 鼻咽肿瘤, 细胞增殖, 肿瘤浸润, 细胞运动

Abstract: ObjectiveTo study the effects of osthole on the proliferation, invasion and migration of nasopharyngeal carcinoma cells CNE2, and to investigate the possible molecular mechanism involved in epithelial to mesenchymal transition (EMT) of CNE2. MethodsCNE2 cells were cultured in vitro and were treated with 0, 20, 40 and 80 μg/ml osthole for 24 or 48 hours, and then methyl thiazolyl tetrazolium (MTT) assay and Transwell assay were used to explore their effects on the cell proliferation, invasion and migration while cells treated with 0 μg/ml osthole were used as the control group. Meanwhile, the mRNA and protein levels of markers of EMT (Ecadherin and vimentin ) and Wnt/βcatenin signaling (βcatenin and cyclin D1) were detected by reverse transcriptionpolymerase chain reaction (RTPCR) and Western blotting respectively. ResultsAfter treatment for 24 and 48 hours, the inhibitory rates of treatment with various concentration of osthole (0, 20, 40, 80 μg/ml) were 0.00%±0.00%, 7.45%±0.87%, 14.12%±2.29%, 27.26%±0.43% and 0.00%±0.00%, 13.44%±0.84%, 29.03%±0.78%, 57.49%±1.70%, with significant differences (F=174.33, P<0.001; F=1 041.40, P<0.001), and the following contrast between each two groups met the statistical significance (all P＜0.01). The migration cells per field of CNE2 cells treated with 0, 20, 40, 80 μg/ml osthole for 48 hours were 52.13±4.49, 29.00±4.49, 18.50±1.93, 13.75±2.77, which exhibited a significant difference (F=200.37, P＜0.001), and the following contrast between each two groups met the statistical significance (all P＜0.01). The invasion cells per field of CNE2 cells treated with 0, 20, 40, 80 μg/ml osthole for 48 hours were 46.63±2.87, 24.13±2.87, 16.75±5.29, 11.00±1.77 respectively, which exhibited a significant difference (F=131.92, P＜0.001), and the following contrast between each two groups met the statistical significance (all P＜0.01). Meanwhile, the relative mRNA and protein expressions of Ecadherin in 0, 20, 40 and 80 μg/ml osthole treatedcells (exposure for 48 hours) were 1.00±0.13, 2.61±0.03, 3.12±0.09, 3.60±0.06 (F=20.92, P<0.001) and 0.22±0.03, 0.35±0.01, 0.60±0.04, 0.82±0.03 (F=178.63, P＜0.001) respectively, and the differences were statistically significant, and further pairwise comparison showed the differences were statistically significant (all P＜0.05). Furthermore, the relative mRNA and protein levels of vimentin, βcatenin, cyclin D1 in 0, 20, 40 and 80 μg/ml osthole treatment for 48 hours were statistically significant difference (mRNA level of vimentin: 1.00±0.12, 0.68±0.03, 0.56±0.01, 0.40±0.09, F=9.48, P<0.010; mRNA level of βcatenin: 1.00±0.14, 0.78±0.04, 0.69±0.07, 0.46±0.12, F=4.84, P<0.050; mRNA level of cyclin D1: 1.00±0.09, 0.82±0.03, 0.58±0.09, 0.40±0.03, F=9.49, P<0.010; protein level of vimentin: 0.85±0.02, 0.74±0.01, 0.34±0.01, 0.27±0.01, F=610.58, P<0.001; protein level of βcatenin: 0.83±0.00, 0.44±0.02, 0.39±0.00, 0.23±0.03, F=985.74, P＜0.001; protein level of cyclin D1: 0.86±0.02, 0.67±0.00, 0.35±0.01, 0.25±0.01, F=910.57, P＜0.001), and further pairwise comparison showed the differences were statistically significant (all P＜0.05). ConclusionOsthole can inhibit the proliferation, invasion and migration of CNE2 cells, which is related to the regulation of Wnt/βcatenin signal pathway and then suppressing of EMT.

Key words: Osthole, Nasopharyngeal neoplasms, Cell proliferation, Neoplasm invasiveness, Cell movement