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

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