Musashi-1和尾型同源盒基因2在胃黏膜肠上皮化生中的表达及意义

Expression and significance of musashi-1 and caudal-related homeodomain transcription 2 in gastric mucosa intestinal metaplasia

目的探讨Musashi-1（Msi-1）和尾型同源盒基因2（CDX2）在不同形态胃窦黏膜肠上皮化生中的表达。方法采用回顾性队列研究方法。收集2009年3月至2013年6月郑州大学第二附属医院收治256例胃炎[慢性非萎缩性胃炎（CNAG）46例、萎缩性胃炎伴肠上皮化生210例]患者的临床病理资料。标本取材由2名内镜科医师负责完成，标本送检，由3名病理科医师独立阅片进行病理学诊断。对萎缩性胃炎伴肠上皮化生标本进行胃镜下分型（局灶隆起型、疣状隆起型、“棉絮状”增厚或凹陷型）和黏液染色检查分型（Ⅰ、Ⅱ、Ⅲ型）；对标本进行免疫组织化学染色检测和免疫荧光双标染色检测。计数资料比较采用，检验。正态分布的计量资料以x±s表示，组间比较采用单因素方差分析，两两比较采用LSD检验。结果（1）临床检查结果：胃镜分型：210例萎缩性胃炎伴肠上皮化生患者中，局灶隆起型65例，疣状隆起型85例，“棉絮状”增厚或凹陷型60例。黏液染色分型：Ⅰ型肠上皮化生70例，Ⅱ型肠上皮化生75例，Ⅲ型肠上皮化生65例。（2）免疫组织化学染色检测结果：204例患者的标本（CNAG36例、局灶隆起型萎缩性胃炎伴肠上皮化生52例、疣状隆起型萎缩性胃炎伴肠上皮化生68例、“棉絮状”增厚或凹陷型萎缩性胃炎伴肠上皮化生48例，Ⅰ、Ⅱ、Ⅲ型肠上皮化生分别为56、60、52例）进行免疫组织化学染色检测。在CNAG组织的腺体颈、峡部发现Msi-1少量阳性表达，在萎缩性胃炎伴肠上皮化生组织中表达呈阳性。Msi-1在CNAG、局灶隆起型、疣状隆起型、“棉絮状”增厚或凹陷型萎缩性胃炎伴肠上皮化生标本中阳性表达率分别为25．0％（9／36）、69．2％（36／52）、76．5％（52／68）、75．0％（36／48），几者比较，差异有统计学意义（χ^2=31．850，P〈0．05）。局灶隆起型、疣状隆起型、“棉絮状”增厚或凹陷型萎缩性胃炎伴肠上皮化生Msi—1阳性表达率分别与CNAG比较，差异均有统计学意义（χ^2=16．655，25．714，20．677，P〈0．05）。Msi-1在Ⅰ、Ⅱ、Ⅲ型肠上皮化生组织中阳性表达率分别为71．4％（40／56）、73．3％（44／60）、76．9％（40／52），3者比较，差异无统计学意义（χ^2=0．432，P〉0．05）。CNAG组织中未发现在特定部位CDX2表达增强；在萎缩性胃炎伴肠上皮化生组织的腺体表达明显。CDX2在CNAG、局灶隆起型、疣状隆起型、“棉絮状”增厚或凹陷型萎缩性胃炎伴肠上皮化生组织中阳性表达率分别为13．9％（5／36）、80．8％（42／52）、82．4％（56／68）、83．3％（40／48），几者比较，差异有统计学意义（，=65．973，P〈0．05）。局灶隆起型、疣状隆起型、“棉絮状”增厚或凹陷型萎缩性胃炎伴肠上皮化生CDX2阳性表达率分别与CNAG比较，差异均有统计学意义（χ^2=38．289．45．496，39．886，P〈0．05）。CDX2在Ⅰ、Ⅱ、Ⅲ型肠上皮化生组织中阳性表达率分别为89．3％（50／56）、80．0％（48／60）、76．9％（40／52），3者比较，差异无统计学意义（χ^2=3．102，P〉0．05）。（3）免疫荧光双标染色检测结果：52例患者的标本（CNAG10例、局灶隆起型萎缩性胃炎伴肠上皮化生13例、疣状隆起型萎缩性胃炎伴肠上皮化生17例、“棉絮状”增厚或凹陷型萎缩性胃炎伴肠上皮化生12例）进行免疫荧光双标染色检测。CNAG组织未发现Msi-1和CDX2双阳性共表达细胞。萎缩性胃炎伴肠上皮化生组织中可见杯状细胞周围Msi-1和CDX2双阳性共表达细胞明显增多。局灶隆起型、疣状隆起型、“棉絮状”增厚或凹陷型萎缩性胃炎伴肠上皮化生中Msi-1和CDX2双阳性共表达率分别为41．4％±11．9％、43．7％±12．7％、42．8％±12．7％，3者比较，差异无统计学意义（F=0．131，P〉0．05）。Msi--1和CDX2双阳性共表达在Ⅰ、Ⅱ、Ⅲ型肠上皮化生组织中阳性表达率分别为33．8％±10．6％、43．8％±10．1％、51．0％±2．7％，3者比较，差异有统计学意义（F=9．817，P〈0．05）；Ⅰ型分别与Ⅱ、Ⅲ型比较，差异均有统计学意义（P〈0．05）；而Ⅱ型与Ⅲ型比较，差异无统计学意义（P〉0．05）。结论Msi-1、CDX2、Msi-1和CDX2双阳性的表达与肠上皮化生的3种形态无关；Msi-1和CDX2双阳性共表达可能是肠上皮化生组织向胃黏膜瘤变过程的重要机制之一。

Objective To investigate the expressions between musashi-1 （Msi-1 ） and caudal-related homeodomain transcription 2 （CDX2） in gastric mucosa intestinal metaplasia. Methods The retrospective cohort study was adopted. The clinicopathological data of 256 patients with gastritis [ 46 of chronic non-atrophic gastritis （CNAG） and 210 of atrophic gastritis and intestinal metaplasia] who were admitted to the Second Affiliated Hospital of Zhengzhou University from March 2009 to June 2013 were collected. The specimens were extracted by 2 endoscopy doctors and then were submitted for inspections. Pathological diagnosis was performed by independent film-reading of 3 pathologists. Endoscopy classification was conducted on the specimens of atrophic gastritis and intestinal metaplasia （ focal uplift type, verrucous uplift type, flocculence incrassation or depression type）. Mucus staining examination was also conducted on these for classification （ Type Ⅰ , Ⅱ and Ⅲ）. The specimens were tested by immunohistochemistry and immunofluoreseence double staining tests. Tile comparison of count data was analyzed using chi-square test. Measurement data with normal distribution were presented as x ＋ s. Comparison among groups was done using the one-way variance analysis and pairwise comparison was done by the LSD test. Results （ 1 ） Results of gastroscopy classification： among the 210 patients with atrophic gastritis and intestinal metaplasia, there were 65 of focal uplift type, 85 of verrucous uplift type and 60 flocculence incrassation or depression type. Classification of mucus staining examination： among the 210 specimens in patients with atrophic gastritis, type was detected in 70 specimens, type Ⅱ in 75 specimens and type Ⅲ in 65 specimens. （2） The 204 specimens were examined by immunobistochemistry test, including 36 of CNAG and 168 of atrophic gastritis and intestinal metaplasia （52 of focal uplift type, 68 of verrucous uplift type and 48 of flocculence incrassation or depression type; 56 of type Ⅰ, 60 of type Ⅱ and 52 of type Ⅲ）. A few expressions of Msi- 1 were detected in the tissues of glandular neck or isthmus in patients with CNAG, and positive expression of Msi- 1 was also detected in the tissues of patients with atrophic gastritis and intestinal metaplasia. The positive expression rates of Msi- 1 in the tissues of patients with CNAG and with atrophic gastritis and intestinal metaplasia of focal uplift type, verrucous uplift type, floeeulence incrassation or depression type were 25.0% （9/36） , 69.2% （36/52） , 76.5% （52/68） and 75.0% （36/48） , respectively, with a significant difference among them （χ^2=31. 850, P 〈0.05） , and with significant differences between CNAG and with atrophic gastritis and intestinal metaplasia of focal uplift type, verrucous uplift type, floeeulence inerassation or depression type （χ^2 = 16. 655,25.714, 20. 677, P 〈 0.05 ）. The positive expression rates of Msi-1 in the tissues of patients with type Ⅰ, type Ⅱ and type Ⅲ of intestinal metaplasia were 71. 4% （40/56）, 73. 3% （44/60） and 76. 9% （40/52） , respectively, with no significant difference among them （χ^2= 0. 432, P 〉 0.05）. There were no increased expressions of CDX2 in the tissues of patients with CNAG and obvious expressions of CDX2 in tissues of the gland of patients with atrophic gastritis and intestinal metaplasia. The positive expression rates of CDX2 in the tissues of patients with CNAG and with atrophic gastritis and intestinal metaplasia of focal uplift type, verrueous uplift type, floceulence incrassation or depression type were 13.9% （5/36）, 80.8% （42/52） , 82.4% （56/68） and 83.3% （40/48） , respectively, with significant difference among them （χ^2= 65. 973, P 〈 0.05 ） , and showing significant differences between focal uplift type, verrucous uplift type, flocculence incrassation or depression type of intestinal metaplasia and CNAG （χ^2= 38. 289, 45. 496, 39. 886, P〈0.05）. The positive expression rates of CDX2 in the tissues with type Ⅰ, type Ⅱ and type Ⅲ of intestinal metaplasia were 89.3% （50/56） , 80.0% （48/60） and 76.9% （40/52） , respectively, with no significant difference among them （χ^2= 3. 102, P 〉 0.05 ）. （3） Results of immunofluoreseence double staining test： the specimens of 52 patients [ 10 of CNAG and 42 of atrophic gastritis and intestinal metaplasia （ 13 of focal uplift type, 17 of verrucous uplift type and 12 of floeeulenee incrassation or depression type）] were detected by immunofluorescence double staining. No double positive co-expression cell was detected in the tissues of patients with CNAG. There was a significant increase of double positive co-expression cells around goblet cells in the tissues of patients with atrophic gastritis and intestinal metaplasia. The co-expression rates of Msi-1 and CDX2 in the double positive cells of patients with focal uplift type, verrueous uplift type, floeeulenee inerassation or depression type were 41.4%±11.9% , 43.7%±12.7% and 42.8%±12.7% , respectively, with no significant difference among them （ F =0. 131, P 〉0.05）. The positive co-expression rates of Msi-1 and CDX2 in the double positive cells of patients with type Ⅰ, type Ⅱ and type Ⅲ of intestinal metaplasia were respectively 33.8% ± 10.6% , 43.8% ±10.1% and 51.0% ± 2.7% , with significant differences among them （ F = 9. 817, P 〈 0.05） and between type Ⅰ and type Ⅱ or type Ⅲ（ P 〈 0.05） , and with no significant difference between type Ⅱ and type m （P 〉 0. 05 ）. Conclusion Expressions of Msi- 1 and CDX2 are not related to the different 3 types of intestinal metaplasia, and co-expression of Msi-1 and CDX2 might be one of the important mechanisms during the transformation from intestinal metaplasia to gastric mucous neoplasms.