用PCR-SSCP方法研究子宫颈癌组织中人乳头瘤病毒E_7基因的变异

USING PCR-SSCP METHOD TO DETECT THE ABNORMALITY OF HPV16 E_7 GENE IN THE CERVICAL CANCER TISSUES

运用核酸杂交和聚合酶链式反应（ＰＣＲ）对临床确诊的６１份子宫颈癌Ⅱ～Ⅲ期标本进行了人乳头瘤病毒（ＨＰＶ）１６型Ｅ７基因的检测。非同位素地高辛标记探针斑点杂交阳性检出率为４５．９％，ＰＣＲ检测阳性率为５４．１％，两种检测结果证实ＨＰＶ－１６型感染与子宫颈癌的发生密切相关。进一步用灵敏的单链构象多态性分析法（ＳＳＣＰ）对３３例阳性标本进行分析，发现有２７例单链ＤＮＡ迁移率与正常对照有不同，推测均发生了ＤＮＡ序列的改变，提示ＨＰＶ－１６Ｅ７基因的变异在子宫颈癌组织中是较常见的现象，其与子宫颈癌的发生有一定的关系。

in this research, digoxigenin labeledprobes of HPV-16 E7 gene were used in dot blot hy-bridization and a set of primers specific to HPV-16Ed gene were used in PCR.(上接第12页) Sixty-one different clinical cervicalcancer specimens were examined with nucleic acidhybridization and PCR. The HPV-16 E_7 gene postive rates were 45. 9% (28/61) and 54. 1% (33/61 )respectively. The results suggested that there was aclose relationship between HPV-16 infection andcervical carcinomas occurence. In order to knowbetter about the abnormality of E_7 gene in the cervical cancer tissues, a sensitive PCR-SSCP methodwas used to detect 33 specimens. Twenty-sevenspecimens had mutations in their single strandDNA, which revealed that the mutations of E_7 genewere the frequent phenomina in the cervical cancers.