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目的 确定在原发性骨肉瘤中是否有ｐ１６ 基因的异常及ｐ１６ 基因异常与ＣＤＫ 基因扩增之间的关系。方法 应用定量Ｓｏｕｔｈｅｒｎ ｂｌｏｔ 方法分析确定ｐ１６ 基因的缺失及ＣＤＫ 基因的扩增。用ＰＣＲＳＳＣＰ（ｐｏｌｙｍｅｒａｓｅｃｈａｉｎ ｒｅａｃｔｉｏｎ ａｎｄｓｉｎｇｌｅｓｔｒａｎｄ ｃｏｎｆｏｒｍａｔｉｏｎ ｐｏｌｙｍｏｒｐｈｉｓｍ） 方法检测ｐ１６ 基因突变。结果 在６０ 例分析ｐ１６ 基因的标本中有５ 例（９％ ）存在ｐ１６ 基因的重排或缺失。均发生于原发肿瘤。在６７ 例分析ＣＤＫ４ 基因的标本中，有６ 例（９％ ）存在ＣＤＫ４ 基因的扩增，包括３ 例原发及３ 例转移的骨肉瘤标本。ｐ１６ 基因异常及ＣＤＫ４ 基因的扩增发生于不同的骨肉瘤标本中，未见相互重叠。在４６ 例同时进行ｐ１６ 及ＣＤＫ４ 基因分析的标本中，２２ ％ 的标本有二者其一的的异常。结论 结合以前的报道，７５ ％ 的骨肉瘤有Ｒｂ 基因的异常提示细胞周期中Ｇ１ 到Ｓ期的异常是骨肉瘤发病机制的特征。
Objective To determine whether there is abnormality of p16 gene and the relationship between p16 gene abnormality and CDK gene amplification in primary osteosarcoma. Methods The quantitative Southern blot analysis was used to determine the deletion of the p16 gene and amplification of the CDK gene. PCR-SSCP (polymerase chain reaction and single-strand conformation polymorphism) was used to detect p16 gene mutations. Results The rearrangement or deletion of the p16 gene was detected in 5 out of 60 specimens analyzed for the p16 gene (9%). All occurred in the primary tumor. Among 67 specimens analyzed for CDK4 gene, 6 (9%) had CDK4 gene amplification, including 3 primary and 3 metastatic osteosarcoma specimens. Abnormality of p16 gene and amplification of CDK4 gene occurred in different osteosarcoma specimens, and no overlap was observed. Of the 46 specimens that were simultaneously analyzed for the p16 and CDK4 genes, 22% had abnormalities in both. Conclusions Combined with previous reports, abnormalities of Rb gene in 75% of osteosarcomas suggest that G1 to S phase abnormalities in the cell cycle are characteristic of the pathogenesis of osteosarcoma.