口腔疾病防治 ›› 2020, Vol. 28 ›› Issue (6): 355-360.DOI: 10.12016/j.issn.2096-1456.2020.06.003

• 基础研究 • 上一篇    下一篇

大蒜素联合5-氟尿嘧啶对黏液表皮样癌MEC-1细胞增殖与凋亡的作用及机制

吴发印(),徐海丽   

  1. 遵义医科大学第五附属(珠海)医院口腔颌面外科,广东 珠海(519100)
  • 收稿日期:2019-09-05 修回日期:2019-12-12 出版日期:2020-06-20 发布日期:2020-05-28
  • 通讯作者: 吴发印
  • 作者简介:徐海丽,医师,硕士,Email: 906760828@qq.com
  • 基金资助:
    贵州省科技技术基金项目(黔科合J字LKZ[2010]07号)

Effect and mechanism of allicin combined with 5-fluorouracil on proliferation and apoptosis of the MEC-1 cell line in mucoepidermoid carcinoma

WU Fayin(),XU Haili   

  1. Department of Oral and Maxillofacial Surgery, the Fifth Affiliated Hospital of Zunyi Medical University, Zhuhai 519100, China
  • Received:2019-09-05 Revised:2019-12-12 Online:2020-06-20 Published:2020-05-28
  • Contact: Fayin WU

摘要:

目的 探讨大蒜素联合5-氟尿嘧啶(5-fluorouracil,5-FU)对黏液表皮样癌MEC-1细胞增殖抑制和凋亡的作用及机制,为临床药物应用提供相应基础。方法 取对数生长期的MEC-1细胞,随机分成对照组和实验组,对照组为含有0.1%DMSO的PBS组,实验组为大蒜素组、5-FU组、联合用药组(大蒜素联合5-FU组)。用CCK8法检测不同浓度(0、25、50、75 mg/L)大蒜素、5-FU及大蒜素联合5-FU作用于MEC-1细胞24 h后细胞增殖抑制率并计算作用24 h后大蒜素及5-FU IC50值。用流式细胞仪检测不同浓度(0、25、50、75 mg/L)大蒜素、5-FU及大蒜素联合5-FU作用于MEC-1细胞24 h后细胞凋亡率。使用Western blot检测IC50浓度的大蒜素、5-FU单独及联合作用于MEC-1细胞24 h后Bax、Bcl-2蛋白表达量。结果 联合用药组对MEC-1的增殖抑制率及凋亡率均强于大蒜素组和5-FU单独用药组(P<0.01)。大蒜素与5-FU单独及联合使用均可以下调Bcl-2蛋白和上调Bax蛋白表达量,且联合用药组Bax/Bcl-2比值最大(P<0.05)。结论 大蒜素和5-FU单独及联合使用均对MEC-1细胞具有增殖抑制作用和诱导凋亡作用,且大蒜素增强了5-FU对MEC-1细胞的凋亡作用,其诱导MEC-1凋亡机制可能与Bax/Bcl-2信号通路相关。

关键词: 黏液表皮样癌, 大蒜素, 5-氟尿嘧啶, 增殖, 凋亡, Bcl-2蛋白, Bax蛋白

Abstract:

Objective To investigate the effect and mechanism of allicin combined with 5-FU on proliferation inhibition and apoptosis of the mucoepidermoid carcinoma MEC-1 cell line in mucoepidermoid carcinoma in order to provide the corresponding basis for subsequent clinical drug application. Methods MEC-1 cells in the logarithmic growth phase were randomly divided into control groups and experimental groups. The control groups were PBS groups containing 0.1% DMSO, while the experimental groups were the allicin group, 5-FU group and combined drug group (the allicin combined with the 5-FU group). The proliferation inhibition rates of allicin, 5-FU and allicin combined with 5-FU in MEC-1 cells were detected by the CCK8 method at different concentrations (0, 25, 50, and 75 mg/L) for 24 h, and the IC50 value of allicin and 5-FU after 24 hours was calculated. The apoptotic rate of MEC-1 cells treated with allicin, 5-FU and allicin combined with 5-FU at different concentrations (0, 25, 50, and 75 mg/L) for 24 hours was measured by flow cytometry. The expression of Bax and Bcl-2 protein was determined by Western blot analysis of the IC50 concentration of allicin and 5-FU alone and in combination with MEC-1 cells for 24 hours. Results The growth inhibition rate and apoptosis rate of MEC-1 cells in the combined drug group were higher than those in the allicin group and the 5-FU alone group (P < 0.01). Allicin and 5-FU alone and in combination downregulated Bcl-2 protein and upregulated Bax protein expression, and the combined drug group had the largest ratio of Bax/Bcl-2 (P < 0.05). Conclusion Allicin and 5-FU both alone and in combination can inhibit the proliferation of and induce apoptosis in MEC-1 cells, and allicin can enhance the apoptosis of 5-FU in MEC-1 cells, which may be related to the apoptosis of the mitochondrial pathway.

Key words: mucoepidermoid carcinoma, allicin, 5-FU, proliferation, apoptosis, Bcl-2 protein, Bax protein

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