白藜芦醇联合NVP-BEZ235对口腔黏膜恶性黑色素瘤抑制作用的实验研究

doi:10.12439/kqhm.1005-4979.2025.06.005

摘要/Abstract

摘要：

目的： 探究白藜芦醇（resveratrol，Res）联合NVP-BEZ235（BEZ235）对口腔黏膜恶性黑色素瘤（oral mucosal malignant melanoma，OMM）细胞系Mm9h-1细胞增殖、侵袭、迁移的影响，并探究其机制。方法： 通过CCK8实验检测药物处理后的细胞存活率，筛选药物浓度并分组，最终确定Control组、100 μmol/L Res组、1.0 μmol/L BEZ235组和联合用药组（C1组：100 μmol/L Res+1.0 μmol/L BEZ235）处理48 h进行后续实验。通过细胞划痕实验和Transwell实验分别评估药物对于Mm9h-1细胞迁移、侵袭能力的影响，Hoechst染色观察细胞死亡形态，蛋白质印迹法检测凋亡相关蛋白的表达水平。结果： Res、BEZ235单药均对Mm9h-1细胞的多种生物学行为有抑制作用。与Control组、100 μmol/L Res组和1.0 μmol/L BEZ235组比较，C1组Mm9h-1细胞存活率、侵袭及迁移能力均显著下降，凋亡相关蛋白的表达显著上调。结论： Res与BEZ235联合用药较单药能更强地抑制Mm9h-1细胞的存活、侵袭和迁移，并能更有效诱导细胞凋亡。

关键词: 白藜芦醇, NVP-BEZ235, 口腔黏膜恶性黑色素瘤, 细胞凋亡

Abstract:

Objective: To investigate the effects of resveratrol (Res) combined with NVP-BEZ235 (BEZ235) on the proliferation, invasion, and migration of the oral mucosal malignant melanoma (OMM) cell line Mm9h-1, and to explore the underlying mechanisms. Methods: Cell viability after drug treatment was detected using the CCK8 assay to determine drug concentrations and group assignments. The following groups were established for 48-hour treatment: Control group, 100 μmol/L Res group, 1.0 μmol/L BEZ235 group, and combination group (C1 group: 100 μmol/L Res+1.0 μmol/L BEZ235). The scratch wound healing assay and Transwell assay were used to evaluate cell migration and invasion abilities, respectively. Hoechst staining was applied to observe cell death morphology, and Western blotting was performed to detect the expression levels of apoptosis-related proteins. Results: Both Res and BEZ235 alone inhibited multiple biological behaviors of Mm9h-1 cells. Compared with the Control, 100 μmol/L Res, and 1.0 μmol/L BEZ235 groups, the C1 group showed significantly reduced cell viability, invasion, and migration capabilities, along with significantly upregulated expression of apoptosis-related proteins. Conclusion: The combination of Res and BEZ235 inhibits Mm9h-1 cell survival, invasion, and migration more effectively than either agent alone, and more potently induces apoptosis.

Key words: resveratrol, NVP-BEZ235, oral mucosal malignant melanoma, cell apoptosis

中图分类号:

R782
R739.8

刘中洋, 杨志恒, 宋宁扬, 李乔乔, 宋继宏, 朴松林. 白藜芦醇联合NVP-BEZ235对口腔黏膜恶性黑色素瘤抑制作用的实验研究[J]. 《口腔颌面外科杂志》, 2025, 35(6): 456-462.

LIU Zhongyang, YANG Zhiheng, SONG Ningyang, LI Qiaoqiao, SONG Jihong, PIAO Songlin. An experimental study on the inhibitory effect of resveratrol combined with NVP-BEZ235 on oral mucosal malignant melanoma[J]. 《Journal of Oral and Maxillofacial Surgery》, 2025, 35(6): 456-462.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2025/V35/I6/456

图/表 5

图1 相同时段内各组药物对Mm9h-1细胞存活率的影响 A.不同药物处理24 h对细胞存活率的影响；B.不同药物处理48 h对细胞存活率的影响；C.各组间吸光度值比较。*表示P<0.05，**表示P<0.01，***表示P<0.001，与Control组比较；#表示P<0.05，##表示P<0.01，与C1组比较。

Figure 1 Effects of different drug groups on the survival rate of Mm9h-1 cells within the same time period

图2 各组药物处理48 h对Mm9h-1细胞迁移能力的影响 A.不同药物处理48 h后细胞划痕实验结果；B.不同药物处理48 h对细胞迁移能力的影响，*表示P<0.05，**表示P<0.01，***表示P<0.001，与Control组比较；#表示P<0.05，##表示P<0.01，与C1组比较。

Figure 2 Effects of different drug groups treatment for 48 h on the migration ability of Mm9h-1 cells

图3 各组药物处理48 h对Mm9h-1细胞侵袭能力的影响 A.药物处理48 h后的细胞Transwell实验结果（×100）；B.药物处理48 h对细胞侵袭能力的影响，***表示P<0.001，与Control组比较；#表示P<0.05，##表示P<0.01，与C1组比较。

Figure 3 Effects of different drug treatments for 48 h on the invasion ability of Mm9h-1 cells

图4 联合用药可能诱导细胞凋亡 A. Hoechst染色法检测各组凋亡细胞（×100）；B. 药物处理细胞后的荧光强度比较，***表示P<0.001，与Control组比较；##表示P<0.01，###表示P<0.001，与C1组比较。

Figure 4 The combination of drugs may potentially induce cell apoptosis

图5 Res联合BEZ235促进Mm9h-1细胞凋亡相关蛋白表达 A.蛋白质印迹法检测凋亡相关蛋白表达；B、C.各组细胞凋亡相关蛋白表达水平比较。*表示P<0.05，**表示P<0.01，***表示P<0.001，与Control组比较；#表示P<0.05，##表示P<0.01，###表示P<0.001，与C1组比较。

Figure 5 The combination of Res and BEZ235 promotes the expression of apoptosis-related proteins in Mm9h-1 cells

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