【摘要】：目的 研究Smad4與β-catenin在兒童腸息肉中的表達(dá)及作用機(jī)制。 方法 選取鄭州大學(xué)第二附屬醫(yī)院病理科2005年7月-2013年4月手術(shù)兒童腸息肉蠟塊標(biāo)本60例，均為幼年性息肉。其中單發(fā)幼年性息肉36例（男性19例，女性17例，年齡3月-12歲，平均年齡6歲±2.1歲），多發(fā)幼年性息肉24例（男性14例，女性10例，年齡6月-9歲，平均4歲±1.3歲）；取成人結(jié)腸腺癌蠟塊15例（男性8例，女性7例，年齡42-83歲，平均56±12.1歲），均為均為管狀腺癌Ⅱ級(jí)；息肉旁正常腸黏膜組織標(biāo)本10例（男性5例，女性5例，年齡5-13歲，平均7±1.2歲），均取自距息肉邊緣大于7cm的正常腸黏膜組織。應(yīng)用免疫組化的方法來測定所選標(biāo)本中的β-catenin與Smad4蛋白的表達(dá)水平。 結(jié)果 β-catenin蛋白表達(dá)情況：正常組中β-catenin蛋白主要為線性表達(dá),蛋白染色為棕黃色顆粒，主要定位于細(xì)胞與細(xì)胞連接處的細(xì)胞膜，陽性率表達(dá)為100%；單發(fā)幼年性息肉組織中的β-catenin蛋白陽性表達(dá)率為80.56%（29/36），少數(shù)標(biāo)本陰性表達(dá)（7例標(biāo)本細(xì)胞漿表達(dá)，2例有胞漿和胞核有弱染色）；多發(fā)幼年性息肉組織中β-catenin染色陽性率為66.67%（16/24）（8例標(biāo)本胞漿染色，3例標(biāo)本胞漿與胞核染色）；大腸腺癌黏膜組織的β-catenin陽性率僅為26.7%（4/15），胞漿和胞核的表達(dá)明顯增加，細(xì)胞膜表達(dá)有不同程度的減少。單發(fā)幼年性息肉組織與多發(fā)幼年性息肉組織中β-catenin蛋白均出現(xiàn)胞質(zhì)和胞核染色。作為陽性對(duì)照的成人結(jié)腸低分化腺癌，β-catenin蛋白為異位表達(dá)。經(jīng)過免疫組化定量分析，與正常對(duì)照組相比，β-catenin蛋白含量在單發(fā)幼年性息肉組、多發(fā)幼年性息肉組、腺癌組均有不同程度的增高，其差異有統(tǒng)計(jì)學(xué)意義。 Smad4蛋白表達(dá)情況:在正常對(duì)照組中Smad4蛋白陽性染色為胞漿或胞核中可見棕色或者黃色顆粒，其陽性率表達(dá)為100%；單發(fā)幼年性息肉組織中的Smad4蛋白染色為陽性，其陽性表達(dá)率為88.89%（32/36）；多發(fā)幼年性息肉組織中Smad4蛋白染色陽性較正常組及單發(fā)幼年性息肉組減弱，其陽性率為70.83%（17/24）；大腸腺癌組織的Smad4呈弱陽性或無表達(dá)，陽性率僅為46.67%（7/15）。單發(fā)幼年性息肉組與腺癌組染色陽性率經(jīng)統(tǒng)計(jì)軟件分析，差異有統(tǒng)計(jì)學(xué)意義；多發(fā)幼年性息肉組與腺癌組染色陽性率比較，差異無統(tǒng)計(jì)學(xué)意義。經(jīng)過免疫組化定量分析，與正常對(duì)照組相比，Smad4蛋白表達(dá)均有不同程度降低。 結(jié)論 本研究所選取的兒童腸息肉標(biāo)本中，不論單發(fā)幼年性息肉還是多發(fā)幼年性息肉，其β-catenin蛋白表達(dá)均較正常對(duì)照組有不同程度的增加，，且其在細(xì)胞的表達(dá)定位發(fā)生改變，由正常的膜線性表達(dá)轉(zhuǎn)化為胞質(zhì)和胞核中有不同程度的染色，提示癌基因β-catenin在兒童腸息肉的形成中起重要作用。而Smad4蛋白均有不同程度表達(dá)減低，且其染色情況有不同程度的減弱，表明抑癌基因Smad4在兒童腸息肉中的作用減弱。二者參與了兒童腸息肉發(fā)生、發(fā)展的過程。
[Abstract]:Objective to study the expression and mechanism of Smad4 and 尾-catenin in intestinal polyps in children. Methods from July 2005 to April 2013, 60 specimens of intestinal polyps were collected from Department of Pathology, second affiliated Hospital of Zhengzhou University, all of them were juvenile polyps. There were 36 cases of single juvenile polyps (male 19 cases, female 17 cases, 3-12 years old, mean age 6 鹵2.1 years), and 24 cases of multiple juvenile polyps (male 14 cases, female 10 cases, 6-9 years old, mean 4 鹵1.3 years old). 15 adult colon adenocarcinoma cases (male 8, female 7; age 42-83 years, mean 56 鹵12. 1 years) were collected, and 10 specimens of normal intestinal mucosa adjacent to polyp were collected (male 5, female 5, age 5-13 years old). The average age was 7 鹵1. 2 years old. All the normal intestinal mucosa tissues were obtained from the margin of the polyp larger than that of 7cm. The expression levels of 尾-catenin and Smad4 protein were determined by immunohistochemical method. Results the expression of 尾-catenin protein: in the normal group, 尾-catenin protein was mainly expressed linearly, and the protein was stained by brown granules, mainly located at the cell membrane at the junction between cell and cell, and the positive rate was 100%. The positive expression rate of 尾-catenin protein was 80.56% (29 / 36) in single juvenile polyps, and a few samples were negative (7 cases expressed in cytoplasm and 2 cases weakly stained in cytoplasm and nucleus). The positive rate of 尾-catenin staining was 66.67% (16 / 24) in multiple juvenile polyps (8 cases with cytoplasm staining, 3 cases with cytoplasm and nucleus staining), and the positive rate of 尾-catenin in colorectal adenocarcinoma mucosa was only 26.7% (4 / 15), the expression of cytoplasm and nucleus increased significantly. Cell membrane expression decreased in varying degrees. Cytoplasmic and nuclear staining of 尾-catenin protein was found in single juvenile polyps and multiple juvenile polyps. 尾-catenin protein was expressed ectopic in adult low differentiated adenocarcinoma of colon as a positive control. Compared with normal control group, 尾-catenin protein content was increased in single juvenile polyp group, multiple juvenile polyp group and adenocarcinoma group. The expression of Smad4 protein: in the normal control group, brown or yellow granules were found in the cytoplasm or nucleus, and the positive rate was 100%. The positive rate of Smad4 protein in single juvenile polyps was 88.89% (32 / 36), and the positive rate of Smad4 protein in multiple juvenile polyps was 70.83% (17 / 24). The positive rate of Smad4 was only 46.67% (7 / 15). The positive rate of staining in single juvenile polyp group and adenocarcinoma group was statistically significant, but there was no significant difference between multiple juvenile polyp group and adenocarcinoma group. Compared with the normal control group, the expression of Smad4 protein decreased in different degree by immunohistochemical quantitative analysis. Conclusion the expression of 尾-catenin protein in children with intestinal polyps was significantly higher than that in normal controls, regardless of single juvenile polyps or multiple juvenile polyps. The expression localization of 尾-catenin was changed from normal membrane linear expression to cytoplasmic and nuclear staining, suggesting that 尾-catenin plays an important role in the formation of intestinal polyps in children. However, the expression of Smad4 protein was decreased in different degree and its staining was weakened in different degree, which indicated that the role of tumor suppressor gene Smad4 in intestinal polyps in children was weakened. Both participate in the process of occurrence and development of intestinal polyps in children.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R735.35

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