Ran Liwei, Tan Weiming, Tan Shengshun, Zhang Ru, Wang Wanjuan, Zeng Weihui
Department of Dermatology and Venereology, the Second Hospital, Xian Jiaotong University, Xian 710004, China.
J Huazhong Univ Sci Technolog Med Sci. 2005;25(4):393-6. doi: 10.1007/BF02828205.
To investigate the effects of ATRA, acitretin and tazarotene on the growth and apoptosis of human tongue squamous cell carcinoma cell line Tca8113. The effect of retinoids on growth of Tca8113 cells in vitro was examined by MTT assay and Trypan blue exclusion assay. Cell cycle analysis, early apoptosis analysis with double staining with Annexin V-FITC and PI, and active caspase-3 analysis with the staining of FITC-conjugated monoclonal rabbit anti-active caspase-3 antibody were made by flow cytometer. Streptavidin-biotin complex (SABC) immunocytochemical assays were employed for the detections of Bax/Bcl-2 proteins expressions. Our results showed that the retinoids inhibited growth of Tca8113 cells in a dose- and time-dependent manner with maximal inhibition 24 h after treatment of 10(-5) mol/L. 10(-5) mol/L retinoids altered cell cycle distribution of Tca8113 cells, revealing an increase in G0/G1-phase population, a decrease in S-phase population and the inhibition of G1/S switching. 10(-5) mol/L retinoids significantly induced apoptosis of Tca8113 cells (all P < 0.05), elevated the cells population with detectable active caspase-3 (P < 0.05 for all), increased the number of cells forming Bax and decreased the number of cells forming Bcl-2 significantly (all P < 0.05). Acitretin played a most prominent role among the retinoids. It is concluded that the inhibition of cell cycle progress of Tca8113 cells by ATRA, acitretin and tazarotene is one of the possible mechanisms for proliferation arrest of Tca8113 cells elicited by the retinoids. The retinoids mediate apoptosis in Tca8113 cells that may be caspase-dependent through mitochondria pathway. High concentration retinoids inhibit growth of Tca8113 cells in vitro by interfering with proliferation and inducing apoptosis of cells. Acitretin may be an alternative medicine for the prevention and treatment of tongue squamous cell carcinoma.
探讨全反式维甲酸（ATRA）、阿维A和他扎罗汀对人舌鳞状细胞癌细胞系Tca8113生长和凋亡的影响。采用MTT法和台盼蓝拒染法检测维甲酸类药物对Tca8113细胞体外生长的影响。通过流式细胞仪进行细胞周期分析、用膜联蛋白V-异硫氰酸荧光素（Annexin V-FITC）和碘化丙啶（PI）双染进行早期凋亡分析以及用异硫氰酸荧光素（FITC）偶联的兔抗活性半胱天冬酶-3单克隆抗体染色进行活性半胱天冬酶-3分析。采用链霉亲和素-生物素复合物（SABC）免疫细胞化学分析法检测Bax/Bcl-2蛋白表达。结果显示，维甲酸类药物以剂量和时间依赖性方式抑制Tca捌13细胞生长，在10^（-5）mol/L处理24 h后抑制作用最强。10^（-5）mol/L维甲酸类药物改变了Tca8113细胞的细胞周期分布，表现为G0/G1期细胞比例增加，S期细胞比例减少，且抑制G1/S期转换。10^（-5）mol/L维甲酸类药物显著诱导Tca8113细胞凋亡（均P<0.05），使可检测到活性半胱天冬酶-3的细胞比例升高（均P<0.05），形成Bax的细胞数量增加，形成Bcl-2的细胞数量显著减少（均P<0.05）。在维甲酸类药物中，阿维A起的作用最为显著。结论：ATRA、阿维A和他扎罗汀抑制Tca8113细胞的细胞周期进程是维甲酸类药物引起Tca8113细胞增殖停滞的可能机制之一。维甲酸类药物介导Tca8113细胞凋亡，可能通过线粒体途径依赖半胱天冬酶。高浓度维甲酸类药物通过干扰细胞增殖和诱导细胞凋亡抑制Tca8113细胞体外生长。阿维A可能是预防和治疗舌鳞状细胞癌的一种替代药物。
