摘要
目的 研究已证实维甲酸受体 -β( Retinoic acid receptor-beta,RARβ)对某些类型的恶性肿瘤细胞有诱导分化和凋亡作用。现观察转导 RARβ基因联合全反式维甲酸 ( all trans retinoic acid,ATRA)对 Tca8113细胞的凋亡作用。方法 先应用脂质体将 RARβ基因导入 Tca8113细胞中 ,然后采用活细胞计数、透射电镜、流式细胞仪等方法观察 ATRA对转基因细胞的凋亡影响和生长抑制作用。结果 转 RARβ基因 Tca8113细胞在 1μmol/LATRA作用 10天后 ,细胞生长抑制率达 90 .2 % ;光镜和透射电镜下观察都可见到典型形态变化的凋亡细胞 ;FCM测定结果表明 ,细胞凋亡率高达 80 .7%。而母本细胞 Tca8113凋亡率小于 2 0 %。结论 RARβ基因可能是 ATRA诱导 Tca8113细胞产生显著凋亡作用的“靶分子”。 RARβ基因联合维甲酸治疗某些口腔鳞癌将是有效和可行的。
Objective Our previous study showed that decreased RARβ gene transcription in oral squamous cell carcinoma cells(Tca8113) was associated with abnormal apoptosis and differentiation of malignant cells. The purpose of this study was to unveil the effect on inducing apoptosis by usuing RARβ gene and all trans retinoic acid(ATRA) in Tca8113 cells.Methods Experiments with Tca8113 cells were performed via transfection of RARβ by lipofectamine. Viable cell count assay was used to investigate the inhibition of cell growth. Flow cytometry and electronic microscopic observation methods were employed for determining the apoptotic activity.Results It was demonstrated that transfection of RARβ gene in Tca8113 cells with its subsequent expression resulted in inhibition of cell growth. The morphological changes of transducting RARβ cells by phase contrast microscope and electronic microscopic observation revealed that ATRA was able to induce apoptosis of RARβ introducting cells. The apoptosis was also evidenced by a quantitative assay for evaluation apoptosis based upon DNA flow cytometry analysis. With the latter method, 80.7% of cells were apoptotic by 10 days following exposure to ATRA at the concentration of 1×10 -6 M. In contrast, ATRA induced apoptosis of Tca8113 cells by less than 20% at 1×10 -6 M concentration.Conclusion These results suggested that RARβ may function as a “molecular target” during ATRA inducing apoptosis in Tca8113 cells. The results also demonstrated that the use of combination of RARβ gene and RA may be an effective therapeutic strategy for patients with oral squamous cell carcinoma.
出处
《口腔颌面外科杂志》
CAS
2000年第3期214-218,共5页
Journal of Oral and Maxillofacial Surgery
基金
国家自然科学基金!396 70 787
上海市教委课题资助!S990 2 0 2
关键词
维甲酸受体-β
鳞状细胞癌
凋亡
基因治疗
Retinoic acid receptor-β
Squamous cell carcinoma
Apoptosis
Genetherapy
