摘要
目的：调查Ｐ１６基因是否与原发性肝细胞性肝癌（ＰＨＣＣ）的发生有关。方法：采用多重ＰＣＲ分析法、单链构象多态性（Ｓｉｎｇｌｅ ｓｔｒａｎｄ ｃｏｎｆｏｒｍ ａｔｉｏｎ ｐｏｌｙｍ ｏｒｐｈｉｓｍ ，ＳＳＣＰ）分析法及以ＰＣＲ为基础的甲基化分析法对２５例原发性肝细胞性肝癌（ＰＨＣＣ）标本及其相应的非肿瘤肝组织标本中Ｐ１６ 基因的缺失、点突变及甲基化情况进行检测。用免疫组织化学法对３５例（包括前述已作基因检测的２５ 例）原发性ＨＣＣ肿瘤标本及其相应的非肿瘤组织标本中Ｐ１６ 蛋白的表达情况进行了调查。结果：２５ 例ＨＣＣ肿瘤标本中，３ 例有Ｐ１６基因缺失，其中２例为纯合子缺失，１例为半合子缺失。另发现１例Ｐ１６基因无缺失者有Ｄ９ｓ１６２缺失。无Ｐ１６基因缺失的２２例肿瘤标本中均未发现有Ｐ１６ 基因点突变存在。肿瘤标本中基因的甲基化发生率为２４％ （６／２５），而全部非肿瘤组织均未出现Ｐ１６基因甲基化。在３５例ＨＣＣ肿瘤标本中发现Ｐ１６蛋白表达缺失１６ 例（４５．７％ ），而全部非瘤组织Ｐ１６ 蛋白均表达阳性。Ｐ１６ 蛋白表达缺失与肿瘤的病理分级有关，分级高者（Ⅲ、Ⅳ级）比分级低者更易发生（Ｐ＜ ０．０１）。在２５例作了Ｐ１６基因检?
Objective: To determine whether P16 is involved in the genesis of hepatocellular carcinoma (PHCC). Methods: Specimens of PHCC and noncarcinomatous liver tissues were collected from 25 cases of PHCC. The deletion of P16 gene exon 1 and exon 2 was identified with comparative multiplex PCR, the point mutation of P16 gene exon 2 was examined with single strand conformational polymorphism (SSCP) and the status of P16 gene methylation was screened with PCR based methylation analysis. In addition, 35 specimens of paraffin embedded PHCC tissue including the surrounding noncarcinomatous liver tissues were examined for P16 protein expression with immunohistochemical analysis. Results: 2 homozygous deletions and 1 hemizygous deletion were found in the 25 PHCC specimens. In the remaining 22 specimens, no point mutation was found. Hypermethylation occurred in 24% (6/25) of the cases and the surrounding noncarcinomatous were always unmethylated at the P16 locus. Loss of P16 protein expression occurred in 45.7% (16/25) of the cases and all the noncarcinomatous liver tissues were P16 staining positive. In the 25 cases of PHCC examined for P16 gene, 3 cases showed P16 gene deletion and 6 cases were found to have methylation of P16 gene. And all the 25 cases were absent for P16 protein expression. Conclusion: Inactivation of P16 gene may play and important role in the genesis of primary PHCC.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
1999年第11期813-817,共5页
Journal of Third Military Medical University
基金
华西医科大学博士点基金
关键词
肝癌
P16基因
聚合酶链反应
免疫组化
基因失活
hepatocellular carcinoma
P16 gene
polymerase chain reaction
immunohistochemistry
