三氧化二砷(As_2O_3)诱导人肺腺癌GLC-82细胞凋亡及分子机制的研究

• 作 者: 邓友平 林晨 张雪艳 陈洁平 肖培根 吴晣
• 机 构: 中国医学科学院中国协和医科大学药用植物研究所!北京100094,中国医学科学院中国协和医科大学肿瘤研究所分子肿瘤学国家重点实验室!北京100021,中国医学科学院中国协和医科大学肿瘤研究所分子肿瘤学国
• 出 处: 癌症
• 发表日期: 1999年05期
• 关键词: 人肺腺癌 As_2O_3 GLC-82 细胞凋亡 细胞系 凋亡峰 抑制生长 基因表达 作用机制 细胞周期

目的： 探讨Ａｓ２ Ｏ３ 对人肺腺癌ＧＬＣ８２ 细胞系抑制生长、诱导凋亡的生物学效应及作用机制。方法：通过ＭＴＴ 还原法检测Ａｓ２ Ｏ３ 对该细胞系生长的影响；用光学显微镜、流式细胞仪、ＤＮＡ 凝胶电泳和细胞凋亡原位检测（ＴＵＮＥＬ） 研究Ａｓ２Ｏ３ 诱导细胞凋亡的情况；用ＲＴＰＣＲ、Ｎｏｒｔｈｅｒｎ ｂｌｏｔ 或Ｗｅｓｔｅｒｎ ｂｌｏｔ 分析Ａｓ２ Ｏ３ 对ｃｍｙｃ、ｐ５３、ｐ１６ 和ｂｃｌＸ 等基因表达的影响。结果：Ａｓ２ Ｏ３ 能抑制ＧＬＣ８２ 细胞的生长。Ａｓ２ Ｏ３ 处理ＧＬＣ８２ 细胞后，光学显微镜下可见到明显的凋亡细胞，细胞周期的Ｇ１ 期前有低于２ 倍体的凋亡峰，ＤＮＡ 凝胶电泳显示出典型的凋亡特征：ＤＮＡ 有规律断裂形成的梯状图谱，蛋白水平的检测表明Ａｓ２ Ｏ３ 可使细胞ｃｍｙｃ 基因表达下降，ｐ１６ 和ｐ５３ 表达升高。结论：Ａｓ２ Ｏ３ 能显著抑制ＧＬＣ８２ 细胞生长、诱导细胞凋亡，并主要通过调节ｃｍｙｃ，ｐ１６ 和ｐ５３ 等基因的表达来实现。

Objective: To investigate the biological effects and mechanisms of As2O3 on growth inhibition and apoptosis in human lung adenocarcinoma GLC82 cell line. METHODS: The effect of As2O3 on the growth of this cell line was examined by MTT reduction method; As2O3-induced apoptosis was studied by light microscopy, flow cytometry, DNA gel electrophoresis, and in situ apoptosis (TUNEL) assay; The effect of As2O3 on the expression of cmyc, p53, p16 and bclX genes was analyzed by RTPCR, Northernern blot or Western blot. Results: As2O3 can inhibit the growth of GLC82 cells. After treatment of GLC-82 cells with As2O3, obvious apoptotic cells were observed under the light microscope. Apoptotic peaks below the diploid were observed before the G1 phase of the cell cycle. DNA gel electrophoresis showed typical characteristics of apoptosis: DNA A stepped pattern formed by regular breaks, the detection of protein levels showed that As2O3 decreased the expression of c-myc gene and increased the expression of p16 and p53. CONCLUSION: As2O3 can significantly inhibit the growth of GLC82 cells and induce apoptosis, and is mainly achieved by regulating the expression of cmyc, p16, and p53 genes.