论文部分内容阅读
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 investigate whether the P16 gene is associated with the development of primary hepatocellular carcinoma (PHCC). METHODS: Twenty-five cases of primary hepatocellular carcinoma (PHCC) were analyzed by multiplex PCR, single strand conformation polymorphism (SSCP) analysis, and PCR-based methylation analysis. P16 gene deletions, point mutations, and methylation were detected in specimens and their corresponding non-tumor liver tissue samples. Immunohistochemistry was used to investigate the expression of P16 protein in primary HCC tumor specimens and their corresponding non-tumor tissue specimens in 35 patients (including the previously described 25 genes tested). Results: Of the 25 HCC tumor specimens, 3 had P16 gene deletions, of which 2 were homozygous and 1 was hemizygous. In addition, it was found that 1 case of P16 gene without deletion had D9s162 deletion. No P16 gene point mutation was found in any of the 22 tumor samples without P16 gene deletion. The methylation rate of genes in tumor samples was 24% (6/25), and all non-tumor tissues did not show methylation of P16 gene. In the 35 HCC tumor specimens, 16 cases (45.7%) of P16 protein were deleted, and all non-tumor tissues were positive for P16 protein. Loss of P16 protein expression was associated with the pathological grade of the tumor. Higher grades (III, IV) were more likely to occur than those with low grade (P < 0.01). P16 gene detection was performed in 25 cases.