Molecular Pathology of Highly Metastatic Rat Osteosarcoma

高转移性大鼠骨肉瘤的分子病理学

• 批准号: 11671449
• 负责人: MII Yoshio
• 金额: $ 2.24万
• 依托单位: Nara Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671449/
• 关键词: Osteosarcoma; Malignant Fibrous Histiocytoma; Rat; p53; p16; p19; TGF-β; Smad

1) We applied wild-type p53 gene transfer to the rat osteosarcoma cells by lipofection, and reconstitution of the p53 inhibits cellular growth in rat osteosarcoma COS1NR cell line, and this growth-suppressive effect is partly due to apoptosis involving bcl-2 gene suppression.2) We investigated the p16 and p19gene expression in rat osteosarcoma cell lines, COS1NR, COS2NR and COS4NR and their subcutaneous tumors and lung metastatic nodules, and rat malignant fibrous histiocytoma cell lines, MFH1NR, MFH2NR and their subcutaneous tumors by RT-PCR.The results showed that there were no differences of the expression of p16 and p19 genes among the all cell lines and tumors. The PCR-SSCP analysis also showed no deletions and point mutations in any genes in any tumors investigated.3) We investigated TGF-β isoforms by Multi-probe RNA protection assay in COS1NR, COS2NR, COS4NR, MFH1NR and MFH2NR.The results showed TGF-β 1to be strongly expressed and no TGF-β 2 and 3 detected in MFHs, while TGF-β 1 and 2 was found to be moderately and 3 weakly expressed in OS cell lines. No changes in TGF-β type II receptor expression, and no mutations of TGF-β type II receptor, Smad2 and 4 genes were detected by RT-PCR and RT-PCRrestricted SSCP anaysis.

1)我们通过脂质体转染将野生型p53基因转移到大鼠骨肉瘤细胞中，p53的重建抑制了大鼠骨肉瘤COS 1 NR细胞系的细胞生长，这种生长抑制作用部分是由于涉及bcl-2基因抑制的细胞凋亡。2）我们研究了大鼠骨肉瘤细胞系COS 1 NR中p16和p19基因的表达，用RT-PCR方法检测大鼠恶性纤维组织细胞瘤细胞系MFH 1 NR、MFH 2NR及其皮下瘤和肺转移瘤中p16、p19基因的表达，结果表明，p16、p19基因在各细胞系和肿瘤中的表达无明显差异。3）采用多探针RNA保护实验对COS 1 NR、COS 2NR、COS 4 NR、MFH 1 NR和MFH 2NR的TGF-β亚型进行了检测，结果显示TGF-β 1在MFH中呈强表达，而TGF-β 2和TGF-β 3在MFH中均未检测到。而TGF-β 1和2在OS细胞系中呈中等表达，3呈弱表达。RT-PCR和RT-PCR限制性SSCP分析均未检测到TGF-β Ⅱ型受体表达的改变，TGF-β Ⅱ型受体、Smad 2和Smad 4基因均未发生突变。

项目成果

Kanya Honoki,et al.: "Possible involvement of bcl-2 suppression in wild type p53 gene dependent-cell growth repression in rat osteosarcoma cells"Toxicologic Pathology. (in press).

Kanya Honoki 等人：“大鼠骨肉瘤细胞中野生型 p53 基因依赖性细胞生长抑制中可能涉及 bcl-2 抑制”毒理学病理学。

Kanya Honoki, et al.: "Possible involvement of bcl-2 suppression in wild type p53 gene dependent-cell growth repression in rat osteosarcoma cells"Toxicologic Pathology. 28. 575-579 (2000)

Kanya Honoki 等人：“大鼠骨肉瘤细胞中野生型 p53 基因依赖性细胞生长抑制中可能涉及 bcl-2 抑制”毒理学病理学。