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目的探索结肠癌组织不同区域肿瘤细胞是否存在增殖能力的差异,且这种区域性差异和多重抑癌基因 p16及其可变读码框架 p14基因的表达情况及其甲基化状态的相关性。方法免疫组化法测定癌块不同区域增殖蛋白 Ki67和 P16、P14的表达差异,激光显微切割技术结合甲基化特异性PCR、巢式逆转录 PCR 观察同一癌块不同区域癌细胞 p16及 p14基因甲基化状态的改变及其 mRNA 的表达情况。结果在42份癌组织标本的中心,增殖蛋白 Ki67均呈强阳性表达,而 P16和 P14蛋白表达缺失;在癌组织标本的边缘,有13份 Ki57表达阳性(30.1%)。不同区域癌细胞的增殖能力差异有统计学意义(P<0.05),且与年龄、性别、Dukes 分期、p14的表达无关,但与 p16的表达显著相关(x~2=25.37.P<0.01)。且 p16基因的甲基化状态和 mRNA 的表达也存在明显的区域性差异(P<0.05)。结论人结肠癌组织中存在肿瘤细胞增殖能力的区域性差异,在癌组织边缘肿瘤细胞侵袭力增强的同时增殖能力下降,癌边缘区 p16基因去甲基化后的再表达可能是这一细胞事件的分子原因。
OBJECTIVE: To explore whether there is a difference in the proliferation ability of tumor cells in different regions of colon cancer tissues, and this regional difference is related to the expression of multiple tumor suppressor gene p16 and its variable reading frame p14 gene and its methylation status. Methods Immunohistochemical method was used to determine the expression differences of proliferating proteins Ki67 and P16 and P14 in different regions of tumor blocks. Laser microdissection combined with methylation-specific PCR and nested RT-PCR was used to observe p16 and p14 in different regions of the same tumor block. Changes in gene methylation status and mRNA expression. RESULTS: In the center of the 42 specimens, the Ki67 protein was strongly positive, while the expression of P16 and P14 protein was missing. At the edge of the specimens, there were 13 Ki57 positive cells (30.1%). There was a significant difference in the proliferation ability of cancer cells in different regions (P<0.05), and was not related to age, gender, Dukes stage, and p14 expression, but was significantly related to the expression of p16 (x~2=25.37.P<0.01). . There was also a significant regional difference in the methylation status and mRNA expression of p16 gene (P<0.05). Conclusion There is a regional difference in tumor cell proliferation ability in human colon cancer tissues. The invasiveness of tumor cells at the edge of cancer tissue increases and the proliferative capacity decreases. The reexpression of p16 gene in the marginal region of the cancer may be a consequence of this cell event. The molecular causes.