摘要：

目的： 观察白藜芦醇对顺铂（cisplatin，DDP）引起的口腔鳞状细胞癌（oral squamous cell carcinoma，OSCC）细胞化疗耐药性的影响，并探讨相关机制。方法： 取对数期CAL-27/DDP细胞，随机分为对照组（常规培养）、白藜芦醇组（加入白藜芦醇200 μmol/L）、激动剂组[加入肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）10 ng/mL]、白藜芦醇联合激动组（加入白藜芦醇200 μmol/L、TNF-α10 ng/mL）。各组均培养48 h用于后续实验。MTT法检测细胞增殖抑制率；双染法检测细胞凋亡率；检测细胞内DDP蓄积与潴留；Western blotting法检测细胞P糖蛋白（P-glycoprotein，P-gp）、拓扑异构酶Ⅱ（topoisomorasesⅡ，TopoⅡ）、核因子-κB（nuclear factor-κB，NF-κB）p65、磷酸化NF-κB（phosphorylated-NF-κB，p-NF-κB）p65、核因子抑制蛋白α（inhibition of NF-κB，IκBα）、磷酸化IκBα（phosphorylated-IκBα，p-IκBα）蛋白表达量。结果： 与对照组比较，白藜芦醇组增殖抑制率、凋亡率、药物蓄积量、药物潴留量、TopoⅡ蛋白表达升高，P-gp、p-NF-κB p65/NF-κB p65、p-IκBα/IκBα蛋白表达量的比值降低（P<0.05），激动剂组增殖抑制率、凋亡率、药物蓄积量、药物潴留量、TopoⅡ蛋白表达降低，P-gp、p-NF-κB p65/NF-κB p65、p-IκBα/IκBα蛋白表达量的比值升高（P<0.05）；与白藜芦醇组比较，白藜芦醇联合激动组增殖抑制率、凋亡率、药物蓄积量、药物潴留量、TopoⅡ蛋白表达降低，P-gp、p-NF-κB p65/NF-κB p65、p-IκBα/IκBα蛋白表达量的比值升高（P<0.05）；与激动剂组比较，白藜芦醇联合激动组增殖抑制率、凋亡率、药物蓄积量、药物潴留量、TopoⅡ蛋白表达升高，P-gp、p-NF-κB p65/NF-κB p65、p-IκBα/IκBα蛋白表达量的比值降低（P<0.05）。结论： 白藜芦醇可降低DDP引起的OSCC细胞化疗耐药性，其作用机制可能与抑制NF-κB通路有关。

关键词: 白藜芦醇, 顺铂, 口腔鳞状细胞癌, 化疗耐药性

Abstract:

Objective: To observe the effect of resveratrol on the chemoresistance of oral squamous cell carcinoma (OSCC) cells induced by cisplatin (DDP), and to explore the related mechanism. Methods: CAL-27/DDP cells in logarithmic phase were taken and randomly divided into control group (conventionally cultured), resveratrol group (added resveratrol 200 μmol/L), agonist group [added tumor necrosis factor-α (TNF-α) 10 ng/mL], resveratrol combined with agonist group (added resveratrol 200 μmol/L, TNF-α 10 ng/mL). All groups were cultured for 48 h for subsequent experiments. The inhibition rate of cell proliferation was detected by MTT method. Double staining was used to detect the apoptosis rate. Intracellular DDP accumulation and retention were examined. Western blotting was used to detect the protein expressions of P-glycoprotein (P-gp), topoisomorases Ⅱ (Topo Ⅱ), nuclear factor-κB (NF-κB) p65, phosphorylated NF-κB (p-NF-κB) p65, inhibition of NF-κBα (IκBα) and phosphorylated IκBα(p-IκBα). Results: Compared with the control group, the proliferation inhibition rate, apoptosis rate, drug accumulation, drug retention, and TopoⅡprotein expression were increased, and the expression of P-gp protein, as well as the ratios of p-NF-κB p65/NF-κB p65, p-IκBα/IκBα protein expression were decreased in the resveratrol group (P<0.05). Also compared with the control group, the proliferation inhibition rate, apoptosis rate, drug accumulation, drug retention, and Topo Ⅱ protein expression were decreased, P-gp protein expression, p-NF-κB p65/NF-κB p65, p-IκBα/IκBα protein expression ratios were increased in the agonist group (P<0.05). Compared with the resveratrol group, the proliferation inhibition rate, apoptosis rate, drug accumulation, drug retention, and TopoⅡprotein expression were decreased, while the expression of P-gp protein, and the ratios of p-NF-κB p65/NF-κB p65, p-IκBα/IκBα protein expression were increased in the resveratrol combined agonist group (P<0.05). Compared with the agonist group, the proliferation inhibition rate, apoptosis rate, drug accumulation, drug retention and TopoⅡprotein expression were increased, while the expression of P-gp protein and the ratios of p-NF-κB p65/NF-κB p65, p-IκBα/IκBα protein expression were decreased in the resveratrol combined with agonist group. Conclusion: Resveratrol can reduce the chemotherapy of cisplatin-induced OSCC cells, and its mechanism may be related to the inhibition of NF-κB pathway.

Key words: resveratrol, cisplatin, oral squamous cell carcinoma, chemotherapy resistance

中图分类号: 

• R782

• R739.8