Quan Hai-Ying, Quan Hai-Ying, Zhou Li-Jia, Li Ang-di, Zhang Ze-Bing
Department of Pathology, Stomatological Hospital, Jilin University. Changchun 130021;Jilin Province, China.E-mail:
Shanghai Kou Qiang Yi Xue. 2015 Feb;24(1):30-6.
To study the role and mechanism of autophagy in chemotherapy of oral squamous cell carcinoma, and provide theoretical evidence to improve chemotherapeutic efficacy of oral squamous cell carcinoma patients.
The cell survival rate changes induced by cisplatin (DDP) and chloroquine (CQ) in CAL-27 cells were assayed by methyl thiazolyl tetrazolium method(MTT). The LC3-II expression level was detected by laser scanning confocal microscope; The apoptotic rate was determined by flow cytometry. SPSS17.0 software package was used for statistical analysis.
MTT results showed that compared with the control group, the cell survival rate reduced with the increasing time of DDP and CQ treatment; The optimal concentration of CAL-27 cells was 5 mg/L after treatment with CQ. IC50 of the CAL-27 cells was 5 mg/L after treatment with DDP; MTT results showed that the cell survival rate of CQ+DDP group was significantly lower than control group, CQ group and DDP group (P<0.05). With the action of CQ and DDP to CAL-27 cells for 48 hours, immunofluorescence results showed that the average fluorescence intensity of DDP group was significantly higher than the other 3 groups (P<0.05), while it was significantly lower in CQ group than the other 3 groups (P<0.05). With the action of CQ and DDP to CAL-27 cells for 48 hours, flow cytometry results showed that the cell apoptosis rate of DDP group and CQ+DDP group were significantly higher than control group and CQ group. The cell apoptosis rate of CQ+DDP group was significantly higher than DDP group (P<0.05). With the action of CQ and DDP to CAL-27 cells for 48 hours, cells in G1 phase of DDP group and CQ+DDP group increased, indicating G1 phase blockage. The cell count in G1 phase of CQ+DDP group was significantly higher than DDP group (P<0.05).
Inhibition of autophagy can enhance the chemotherapeutic sensitivity of DDP in CAL-27 cells. Autophagy in CAL-27 cells is an important mechanism for chemotherapy resistance of oral squamous cell carcinoma. Autophagy inhibitor may have significant potential to be a novel chemotherapeutic sensitizer for oral squamous cell carcinoma.
研究自噬在口腔鳞状细胞癌化疗中的作用及机制，为提高口腔鳞状细胞癌患者化疗疗效提供理论依据。
采用甲基噻唑基四氮唑蓝法（MTT）检测顺铂（DDP）和氯喹（CQ）诱导CAL-27细胞的存活率变化。用激光扫描共聚焦显微镜检测LC3-II表达水平；通过流式细胞术测定凋亡率。采用SPSS17.0软件包进行统计分析。
MTT结果显示，与对照组相比，随着DDP和CQ处理时间的增加，细胞存活率降低；CQ处理后CAL-27细胞的最佳浓度为5mg/L。DDP处理后CAL-27细胞的IC50为5mg/L；MTT结果显示，CQ+DDP组的细胞存活率显著低于对照组、CQ组和DDP组（P<0.05）。CQ和DDP作用于CAL-27细胞48小时后，免疫荧光结果显示，DDP组的平均荧光强度显著高于其他3组（P<0.05），而CQ组显著低于其他3组（P<0.05）。CQ和DDP作用于CAL-27细胞48小时后，流式细胞术结果显示，DDP组和CQ+DDP组的细胞凋亡率显著高于对照组和CQ组。CQ+DDP组的细胞凋亡率显著高于DDP组（P<0.05）。CQ和DDP作用于CAL-27细胞48小时后，DDP组和CQ+DDP组G1期细胞增多，提示G1期阻滞。CQ+DDP组G1期细胞计数显著高于DDP组（P<0.05）。
抑制自噬可增强DDP对CAL-27细胞的化疗敏感性。CAL-27细胞中的自噬是口腔鳞状细胞癌化疗耐药的重要机制。自噬抑制剂可能具有成为口腔鳞状细胞癌新型化疗增敏剂的巨大潜力。
