CAJ | 학술논문

目的通过人雄激素受体基因位点克隆性分析技术对胃癌及其癌前病变进行克隆性分析,探讨胃癌发生发展过程中单克隆发生率的变化趋势及与Ki-67表达之间的关系及其意义.方法肠型胃癌根治标本24例,胃镜活检标本150例.采用激光显微切割技术准确获取病变腺上皮细胞,基因组DNA经甲基化敏感的Hpa Ⅱ限制性内切酶消化后,PCR扩增人雄激素受体基因,采用基因扫描技术对PCR产物进行分析.应用免疫组织化学EnVision二步法检测Ki-67在以上病变组织中的表达,并探讨其与克隆性分析结果的相关性.结果单克隆发生率在胃黏膜肠上皮化生(15.63%,5/32)、低级别上皮内瘤变(22.22%,10/45)、高级别上皮内瘤变(69.44%,25/36)及肠型胃癌(100.0%,20/20)中逐渐增加,除胃黏膜肠上皮化生和低级别上皮内瘤变之间的差异没有统计学意义(P=0.47),其他各组之间差异均有统计学意义(P<0.01).Ki-67的阳性表达率随着病变的发展而不断升高.低级别上皮内瘤变组织中单克隆病例的Ki-67的阳性表达率显著高于多克隆病例(P<0.01),且克隆性与Ki-67的阳性表达率之间存在显著相关性(P<0.01).结论单克隆的发生率及Ki-67的阳性表达率在胃癌发生发展过程中逐渐增加.且单克隆病变的发生与Ki-67的表达存在一定的相关性,两者的联合可用于胃癌的早期诊断及易感性的预测.
목적 통과인웅격소수체기인위점극륭성분석기술대위암급기암전병변진행극륭성분석,탐토위암발생발전과정중단극륭발생솔적변화추세급여Ki-67표체지간적관계급기의의.방법 장형위암근치표본24례,위경활검표본150례.채용격광현미절할기술준학획취병변선상피세포,기인조DNA경갑기화민감적Hpa Ⅱ한제성내절매소화후,PCR확증인웅격소수체기인,채용기인소묘기술대PCR산물진행분석.응용면역조직화학EnVision이보법검측Ki-67재이상병변조직중적표체,병탐토기여극륭성분석결과적상관성.결과 단극륭발생솔재위점막장상피화생(15.63%,5/32)、저급별상피내류변(22.22%,10/45)、고급별상피내류변(69.44%,25/36)급장형위암(100.0%,20/20)중축점증가,제위점막장상피화생화저급별상피내류변지간적차이몰유통계학의의(P=0.47),기타각조지간차이균유통계학의의(P<0.01).Ki-67적양성표체솔수착병변적발전이불단승고.저급별상피내류변조직중단극륭병례적Ki-67적양성표체솔현저고우다극륭병례(P<0.01),차극륭성여Ki-67적양성표체솔지간존재현저상관성(P<0.01).결론 단극륭적발생솔급Ki-67적양성표체솔재위암발생발전과정중축점증가.차단극륭병변적발생여Ki-67적표체존재일정적상관성,량자적연합가용우위암적조기진단급역감성적예측.
Objective To study the clonality of gastric carcinoma and precancerous lesions and its relationship with Ki-67 protein expression. Methods Formalin-fixed paraffin embedded tissues were collected from 174 cases of gastric endoscopic biopsies and surgical removed specimens. The lesional tissues were isolated by Laser Capture Microdissection. Methylation sensitive restriction enzyme (Hpa Ⅱ) digestion and polymerase chain reaction (PCR) were used to detect the clonality at the polymorphic human androgen receptor gene locus on the X chromosome. PCR products were analyzed by capillary electrophoresis using applied Biosystems 3730 DNA Analyzer. In addition, a two-step immunohistochemical staining EnVision method was used to detect the expression of Ki-67 protein. Results The frequency of detection of monoclonality and expression rate of Ki-67 were found increased in a stepwise fashion from gastrointestinal metaplasia, low grade intraepithelial neoplasia, high grade intraepithelial neoplasia to intestinal carcinoma (15. 63% ,5/32; 22.22%, 10/45; 69.44%, 25/36 and 100. 0%, 20/20; respectively). The presence of clonal proliferation was correlated with Ki-67 expression in low grade intraepithelial neoplasia (P<0.01).Conclusions The presence of clonal proliferation and increased Ki-67 are increasingly detected in the lesions along the multi-step gastric carcinogenesis model. Clonal status is associated with the expression rate of Ki-67 to a certain extent, suggesting a combined application of both markers may be useful in assessing early stages of gastric carcinoma.