Choo K B, Chong K Y
Department of Medical Research, Veterans General Hospital, Taipei, Taiwan, Republic of China.
Virology. 1993 Apr;193(2):1042-6. doi: 10.1006/viro.1993.1224.
Previous biochemical and structural studies using established cell lines have led to a hypothesis that the cellular p53 and the retinoblastoma susceptibility (Rb) proteins may play a crucial role in the pathogenesis process of cervical carcinoma. To further test this hypothesis, we have analyzed the structure of the p53 and Rb genes in 38 primary cervical carcinoma samples with known HPV status using the polymerase chain reaction-based single-strand conformation polymorphism (SSCP) approach. All 11 exons as well as 500 bp of the 5'-noncoding region of the p53 gene and exons 13-23 of the Rb gene were analyzed. None of these exons in all the samples analyzed showed band shift in the SSCP gel indicative of the presence of mutation. In the same experiments, however, all known exon mutations as well as previously unreported mutations in the intron sequences of the p53 and Rb genes in the HPV-free C33-A and HT-3 cell lines were detected. The absence of mutation in the p53 gene in all five HPV-negative samples was confirmed by direct sequence analysis. It appears that p53 and Rb mutations are a very rare event in cervical cancer and their occurrence is apparently not strictly correlated with HPV status.
以往利用已建立的细胞系进行的生化和结构研究提出了一种假说,即细胞中的p53蛋白和视网膜母细胞瘤易感(Rb)蛋白可能在宫颈癌的发病过程中起关键作用。为了进一步验证这一假说,我们采用基于聚合酶链反应的单链构象多态性(SSCP)方法,分析了38例已知HPV状态的原发性宫颈癌样本中p53和Rb基因的结构。分析了p53基因的所有11个外显子以及5'-非编码区的500 bp片段,以及Rb基因的外显子13 - 23。在所有分析的样本中,这些外显子在SSCP凝胶中均未显示出指示突变存在的条带迁移。然而,在相同的实验中,在不含HPV的C33 - A和HT - 3细胞系中检测到了p53和Rb基因内含子序列中所有已知的外显子突变以及先前未报道的突变。通过直接序列分析证实了所有五个HPV阴性样本中p53基因均无突变。看来p53和Rb突变在宫颈癌中是非常罕见的事件,它们的发生显然与HPV状态没有严格的相关性。