Saito H, Morizane T, Watanabe T, Kagawa T, Miyaguchi S, Kumagai N, Tsuchimoto K, Tsuchiya M
Department of Internal Medicine, School of Medicine, Keio University, Kanagawa Dental College, Tokyo, Japan.
Keio J Med. 1991 Sep;40(3):139-45. doi: 10.2302/kjm.40.139.
Changes of nucleotide sequences and expressions of cellular oncogenes in human hepatoma cell lines, PLC/PRF/5, HCC-M and HCC-T cells, were examined by Southern and Northern blot analyses. The probes used are DNA fragment of myc, N-, H-, K-ras, fos, fms, raf, erb-A, erb-B, and erb-B2 genes and synthetic oligonucleotides corresponding to the part of N-, H-, K-ras genes. The results are as follows. DNA amplification and rearrangement were not detected in these three human hepatoma cell lines. Point mutations at codons 12, 13, and 61 in N- and K-ras genes were not demonstrated in these cell lines. N-, H-, K-ras and myc transcripts were detected in these three cell lines. However, fos gene transcript was detected only in PLC/PRF/5 and HCC-M cells which were derived from hepatitis B related hepatocellular carcinoma and having integrated hepatitis B virus (HBV) DNA. These data showed that there are no specific proto-oncogene expression into RNA except for myc and ras genes, nor DNA rearrangement in these 3 human hepatoma cell lines with regards to at least 10 different oncogenes examined and suggest the relationship between fos gene expression and integration of HBV DNA in host cell DNA.
通过Southern和Northern印迹分析，检测了人肝癌细胞系PLC/PRF/5、HCC-M和HCC-T细胞中核苷酸序列的变化以及细胞癌基因的表达。所用探针为myc、N-、H-、K-ras、fos、fms、raf、erb-A、erb-B和erb-B2基因的DNA片段以及与N-、H-、K-ras基因部分相对应的合成寡核苷酸。结果如下。在这三种人肝癌细胞系中未检测到DNA扩增和重排。在这些细胞系中未证实N-和K-ras基因第12、13和61位密码子的点突变。在这三种细胞系中检测到了N-、H-、K-ras和myc转录本。然而，仅在源自乙型肝炎相关肝细胞癌且整合有乙型肝炎病毒（HBV）DNA的PLC/PRF/5和HCC-M细胞中检测到fos基因转录本。这些数据表明，就所检测的至少10种不同癌基因而言，在这三种人肝癌细胞系中除了myc和ras基因外，没有特定的原癌基因表达为RNA，也没有DNA重排，并提示fos基因表达与宿主细胞DNA中HBV DNA的整合之间的关系。
