口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (12): 1293-1296.DOI: 10.13701/j.cnki.kqyxyj.2018.12.008

• 口腔肿瘤学研究 • 上一篇    下一篇

毒胡萝卜素诱导舌鳞癌细胞Cal-27凋亡的实验研究

戴巧群,陈文英,史芳萍*,陈瑜露   

  1. 浙江省宁波市鄞州区第二医院口腔科 浙江 宁波 315100
  • 收稿日期:2018-01-26 出版日期:2018-12-28 发布日期:2018-12-27
  • 通讯作者: 史芳萍,E-mail:55017618@qq.com
  • 作者简介:戴巧群(1983~ ),女,浙江宁波人,硕士,主治医师,主要从事天然活性物质用于额面部恶性肿瘤防治。

Experimental Study on Thapsigargin Induced Apoptosis in Tongue Cancer Cell Line Cal-27.

DAI Qiao-qun, CHEN Wen-ying, SHI Fang-ping*, CHEN Yu-lu   

  1. Department of Stomatology, Ningbo Yinzhou No.2 Hospital, Ningbo 315100, China.
  • Received:2018-01-26 Online:2018-12-28 Published:2018-12-27

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摘要： 目的: 研究毒胡萝卜素对人舌鳞状细胞癌CAL-27细胞活力及凋亡的影响。方法: 用不同浓度的毒胡萝卜素处理培养的CAL-27细胞,用噻唑蓝(methyl thiazol tetrazolium,MTT)法测定CAL-27细胞增殖情况,4',6-二脒基-2-苯基吲哚(4',6-diamidino-2-phenylindole,DAPI)染色观察细胞形态学变化,通过膜联蛋白V(Annexin V) -异硫氰酸荧光素(fluoresceine isothiocyanate,FITC)/碘化丙啶(propidium iodide ,PI)双染标记流式细胞术检测细胞凋亡的比例,Western blot检测凋亡相关蛋白半胱氨酰天冬氨酸特异性蛋白酶3(cysteinyl aspartate-specific protease-3,Caspase-3) 等活化情况。结果: 毒胡萝卜素剂量依赖性抑制CAL-27细胞生长,其半数抑制浓度(50% inhibitory concentration,IC50)为15.09μmol/L。DAPI染色结果显示,细胞核变小,染色质皱缩。流式细胞仪检测结果显示,随着毒胡萝卜素剂量的增加,晚期凋亡细胞比例增加。Western blot结果显示,毒胡萝卜素浓度与Caspase-3、Caspase-9活化呈正相关,并影响其他凋亡相关蛋白B细胞淋巴瘤/白血病-2(B-cell lymphoma/Leukemia-2,bcl-2)和 B细胞淋巴瘤/白血病-2相关X蛋白(B cell lymphoma/leukemia-2 associated X protein,bax)表达。结论: 毒胡萝卜素可以显著抑制CAL-27细胞增殖,并可诱导CAL-27细胞凋亡,其作用机制与Caspase-3和Caspase-9活化有关。

关键词: 毒胡萝卜素, 舌癌, 细胞凋亡

Abstract: Objective: To study the effects of thapsigargin on the viability and apoptosis of CAL-27 cells.Methods: CAL-27 cells were treated with different concentrations of thapsigargin and detected by methyl thiazolyl tetrazolium (MTT).After double staining with Annexin V-FITC/PI, the proportion of apoptotic cells was detected by flow cytometry.Western blotting was used to detect the activation of Caspase-3 and other apoptosis-related proteins.Results: Different concentrations of thapsigargin could inhibit the growth of CAL-27 cells in a dose-and dependent manner.The IC50 was 15.09 μmol/L.DAPI staining showed that the nucleus becomes smaller and the chromatin shrinks with increasing doses of thapsigargin.Flow cytometry results showed that the apoptotic cells at late phase were increased.When detecting the apoptosis related proteins, the activation of Caspase-3 and Caspase-9 was shown after treatment with thapsigargin.Conclusion: Thapsigargin can significantly inhibit the proliferation of CAL-27 cells and induce the apoptosis of CAL-27 cells.The mechanism of action is related to the activation of Caspase-3 and Caspase-9.

Key words: Thapsigargin, Oral cancer, Apoptosis