摘要
目的:探讨应用全反式维甲酸处理细胞增殖和分化对人骨肉瘤细胞的影响。方法:实验于2004-08/2005-01在第四军医大学唐都医院解放军骨科中心骨肿瘤研究所完成。以1×10-6mol/L的全反式维甲酸处理人骨肉瘤OS-9901细胞为试验组,对照组细胞不加全反式维甲酸处理。观察维甲酸对骨肉瘤细胞诱导分化向良性细胞发展的作用。①计算细胞增殖抑制率,通过四唑盐检测。②用药后细胞形态的变化,以显微镜观察。③骨肿瘤较为特异性的碱性磷酸酶表达变化,应用碱性磷酸酶染色观察。④用药对骨形成蛋白2表达的影响,表达增高促进骨肿瘤细胞的增殖,应用免疫组织化学染色观察。⑤细胞的变化应用透射电镜观察。⑥计算成瘤率,并对试验组与对照组肿瘤体积大小进行比较,采用裸鼠移植瘤试验。结果:①全反式维甲酸处理第2天对人骨肉瘤OS-9901细胞生长的平均抑制率为25.2%,第4天达到51.7%,第6天为68.4%。②显微镜下可见处理后的细胞生长及形态有明显改变,增殖缓慢,生长差,数量减少,分裂像少见。③碱性磷酸酶和骨形成蛋白2的表达减少。④透射电镜下出现处理后细胞膜绒毛减少,胞核光滑无切迹,核浆比例下降等良性分化改变。⑤各组肿瘤的成瘤率均为100%,成瘤期对照组约为7d,实验组约为10d,用药组瘤细胞核浆比例较对照组减小,出现大量的凋亡细胞和大片的坏死区,显示有明显抑制增值和向良性转变倾向。结论:全反式维甲酸有抑制骨肿瘤细胞增殖,并诱导其向良性分化作用。
AIM: To study the effect of cellular proliferation and differentiation with all-trans retinoic acid(ATRA) on human osteogenic sarcoma cell. METHODS: The experiment was completed at the Institute of Orthopedic Oncology, Center of Orthopaedics, Tangdu Hospital, Fourth Military Medical University of Chinese PLA from August 2004 to January 2005. Human osteogenic sarcoma cell line OS-9901 was treated with 1×10-6 mol/L ATRA as the experimental group, and cells in the control group was treated without ATRA in order to observe the effect of ATRA on the differentiation induced by osteogenic sarcoma cell developing into benign cell. ①Inhibitory rate of cell proliferation was calculated with MTT assay. ②Changes of cellular form was observed with microscope after medication. ③Changes of alkaline phosphatase expression in specificity of bone tumor was observed with alkaline phosphatase stain. ④Effect of medication on the expression of bone formatting protein 2 and the effect of increasing expression on the improvement of proliferation of bone tumor cell were observed with immunohistochemical stain. ⑤Cellular changes was observed with transmission electron microscopy. ⑥The rate of neoplasia was calculated and compared with the size of tumor in the experimental group and the control group. The transplantation of tumor was performed on the nude mouse. RESULTS: ①Two days after ATRA treatment, average inhibitory rate of human osteogenic sarcoma cell line OS-9901 was 25.2%, 51.7%on the fourth day, and 68.4%on the sixth day.②The changes of cellular growth and form were observed under the microscope, including slowing proliferation, poor growth, quantity reduction and few disintegration. ③Expressions of alkaline phosphatase and bone formatting protein 2 were decreased.④Changes of benign differentiation was observed under the transmission electron microscopy, including reduction of cell membrane villus, smooth cellular nucleus without incisure, and the descent of karyoplasms ratio.⑤All rate of neoplasis was 100%in each group, and the period of neoplasia was 7 days in the control group and 10 days in the experimental group. The ratio of tumor-nucleus laryoplasm was lower in medication group than that in control group, and massive apoptosis and necrosis were seen, which meant that proliferation was inhibited obviously and developed into benign cell gradually. CONCLUSION: ATRA can inhibit the proliferation of osteogenic sarcoma cells and lead to benign differentiation.
出处
《中国临床康复》
CSCD
北大核心
2005年第22期150-151,i005,共3页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助(30330610)~~
