正常的和转化的C3H小鼠胚胎成纤维细胞neu基因结构的初步研究

应用ＰＣＲ－ＳＳＣＰ技术并结合Ｓｏｕｔｈｅｒｎ印迹杂交从基因水平对正常和￣３Ｈ－ＴｄＲ恶性转化小鼠胚胎成纤维细胞ＮＣ３Ｈ１０和ＴＣ３Ｈ１０中ｎｅｕ基因进行研究。Ｓｏｕｔｈｅｒｎ印迹杂交结果表明恶性转化的ＴＣ３Ｈ１０细胞ｎｅｕ基因出现重排和扩增，ＳＳＣＰ分析未发现ＴＣ３Ｈ１０细胞ｎｅｕ基因跨膜区突变。上述结果说明ＴＣ３Ｈ１０细胞ｎｅｕ基因结构异常可能在跨膜区外，ｎｅｕ基因异常在￣３Ｈ－ＴｄＲ诱导的细胞恶性转化过程中可能有重要的作用，ＥＧＦ可促进ｎｅｕ基因表达增高，研究发现在ＥＧＦ持续作用下，ＮＣ３Ｈ１０细胞ｎｅｕ基因甲基化水平无显著变化，说明ＥＧＦ可能是通过其它途径调控ｎｅｕ基因表达增高的，排除了ＥＧＦ通过改变ｎｅｕ基因甲基化水平而调控ｎｅｕ基因表达的可能性。

Primary Analyses of the Structure of neu Oncogene in Normal and Transformed C3H Mouse Embryo Fibroblasts

The neu oncogene in normal C3H mouse embryo fibroblasts C3H10T1/T2Cl 8 (designated NC3H10 ) and ￣3H-TdR transformed C3H10T1/2Cl 8 (designated TC3H10) were detected by PCR-SSCP and Southern analyses. Rearrangement and amplification of neu oncogene occurred in TC3H10. No point mutation was found in transmembrane domain of neu gene in TC3H10 by PCR-SSCP. These results suggested that aberration of structure of neu gene had not occurred at transmembrane domain. Abnormal of neu oncogene may play an important role in development of ￣3H-TdR transformation. EGF increased expression of neu oncogene. Treatment by EGF led to no obvious change of DNA methylation level of neu gene in NC3H10. though it led to a decrease of DNA methylation level of c-fos. This suggested that effects of EGF on enhancement of neu expression may be controlled by other ways,rather than by decrease of neu DNA methylation level.