Journal of Prevention and Treatment for Stomatological Diseases ›› 2020, Vol. 28 ›› Issue (6): 355-360.doi: 10.12016/j.issn.2096-1456.2020.06.003

• Basic Study • Previous Articles     Next Articles

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 E-mail:WuFY0515@163.com

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

CLC Number: 

  • R78

Table 1

Growth inhibition rate of MEC-1 cells 24 hours after treatment x ± s,n=5,%"

Groups 0 mg/L 25 mg/L 50 mg/L 75 mg/L F P
Allicin group 0 4.692 ± 0.795 16.889 ± 0.9281) 28.861 ± 1.3731)2) 648.490 0.001
5-FU group 0 5.133 ± 0.422 17.011 ± 2.2481) 27.455 ± 1.4191)2) 258.275 0.001
Combined drug group 0 15.874 ± 1.0323)4) 25.311 ± 1.2471)3)4) 38.768 ± 4.0931)2)3)4) 102.483 0.001

Table 2

Apoptosis rate of MEC-1 cells after 24 h of treatment x ± s,n=5,%"

Groups 0 mg/L 25 mg/L 50 mg/L 75 mg/L F P
Allicin group 9.387 ± 0.443 10.933 ± 0.7941) 14.040 ± 0.5051)2) 16.963 ± 1.0091)3) 107.840 0.001
5-FU group 9.387 ± 0.443 11.423 ± 0.3161) 13.120 ± 0.6071)2) 15.770 ± 1.0291)3) 84.804 0.001
Combined drug group 9.387 ± 0.4434)5) 13.020 ± 0.2041)4)5) 17.397 ± 1.4851)2)4)5) 22.383 ± 1.3351)3)4)5) 149.104 0.001

Figure 1

Apoptosis rate of MEC-1 cells after 24 h of treatment different drug groups with 25 mg/L a: control group; b:allicin group, compared with the control group, the apoptosis rate increased; c: 5-FU group, compared with the control group, the apoptosis rate increased; d: combined drug group,compared with other groups, the apoptosis rate increased"

Table 3

Ratio of Bax and Bcl-2 protein expression and Bax/Bcl-2 expression in MEC-1 cells x ± s,n=3,%"

Group Bcl-2 Bax Bax/Bcl-2
Control group 0.907 ± 0.015 0.353 ± 0.153 0.390 ± 0.023
Allicin group 0.580 ± 0.0261) 1.027 ± 0.0211) 1.773 ± 0.0891)
5-FU group 0.567 ± 0.0311) 1.040 ± 0.0441) 1.839 ± 0.1261)
Combined
drug group
0.343 ± 0.0251)2)3) 1.430 ± 0.0401)2)3) 4.182 ± 0.3731)2)3)
F 258.289 575.773 -
P 0.001 0.001 0.022

Figure 2

Expression of Bcl-2 and Bax protein by using allicin and 5-FU alone or in combination 1: allicin group; 2: 5-FU group; 3: combination group; 4: control group"

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