腺样囊腺癌的血管生成拟态的研究及与肿瘤干细胞标志蛋白相关性分析

发布时间：2018-04-14 15:40

本文选题：眼眶 + 泪腺肿瘤　；参考：《天津医科大学》2011年硕士论文

【摘要】：目的 1.本实验在人类腺样囊腺癌组织学标本中寻找并确认血管生成拟态结构,明确其与病理学分型的关系及其表达的意义。 2.观察被肿瘤干细胞标记相关蛋白CD44v6,CD133,ABCG2标记的标本在人类腺样囊腺癌组织学标本中的表达,分析比较血管生成拟态与其之间的相关性,为该肿瘤易于局部复发提供新的解释。 3.从细胞水平再次进行形态学观察,明确血管生成拟态结构在人涎腺腺样囊性癌细胞系SACC-83中存在,并观察肿瘤干细胞标记相关CD44v6,CD133,ABCG2在人涎腺腺样囊性癌细胞系SACC-83中的表达。为进一步肿瘤干细胞的提取及分析VM生成的机制打下基础。方法 1.本实验收集天津医科大学第二医院1999年1月-2009年3月间手术切除的符合实验条件的泪腺腺样囊腺癌组织标本33例。将所有标本统一进行重新切片经HE染色后观察确认病理诊断及分型。通过CD31/PAS双重染色后在光学显微镜下对比观察切片,在人泪腺腺样囊腺癌组织标本中寻找并确认VM结构。同时应用二步法免疫组织化学检测33例标本中CD44v6、CD133和ABCG2的表达情况。 2.根据VM结构在泪腺腺样囊腺癌中的观察情况,应用统计学方法比较其在不同病理分型中的表达以及在VM表达组与非表达组肿瘤干细胞标记相关蛋白CD44v6, CD133, ABCG2的表达有无差异。 3.通过人涎腺腺样囊性癌SACC-83细胞系由北京医科大学口腔医院李盛霖中心实验室馈赠)二维培养及三维培养,观察血管生成拟态及肿瘤干细胞标记相关蛋白CD44v6, CD133, ABCG2在细胞系SACC-83中的表达。结果 1.33例标本经观察HE染色切片,并结合免疫组织化学结果确诊为泪腺腺样囊腺癌,其中实体型13例(39.39%),腺样-管状型20例(60.61%)。通过CD31/PAS双重染色方法观察,在10例标本中发现了VM结构,出现率为30.3%。,实体型6例(46.2%),腺样-管状型4例(20.0%),二者差异不具有统计学意义(p=0.346)。 2.所观察到的VM均由腺样囊腺癌细胞围绕形成管状、裂隙状结构,该腔隙结构内可见红细胞,这些红细胞结构完整,且周围没有出血灶存在。此外还观察到部分瘤细胞胞浆内分泌大量PAS阳性物质及CD31阳性物质。在VM表达组与非表达组CD44v6的表达分别为1.57%与2.51%,二者具有统计学差异(p=0.023)。 3.33例标本应用免疫组织化学的方法观察肿瘤干细胞相关标志蛋白CD44v6,CD133, ABCG2的表达情况分别为：CD44v6阳性表达率为54.5%(18/33)。实体型阳性率76.9%(10/13),腺样-管状型阳性率为40.0%(8/20),二者差异具有统计学意义(p=0.037)。CD133阳性表达率为57.6%(19/33),在实体型和腺样-管状型中分别为76.92%(10/13)、45%(9/20),差异不具有统计学意义(p=0.070)；表达产物定位于细胞膜和细胞浆,呈淡黄色至棕褐色,部分病例同时表达于胞浆和胞核。ABCG2阳性表达率为21.2%(7/33),在实体型和腺样-管状型中分别为30.8%(4/13)、15.0%(3/20),差异不具有统计学意义(p=0.297);ABCG2阳性表达细胞具有沿血管分布的倾向。 4.在涎腺腺样囊性癌细胞系SACC-83的二维培养中,可见细胞呈环状,网状生长,三维培养中呈腔状、管状生长。在三维培养中可见VM阳性表达。肿瘤干细胞相关蛋白在人涎腺腺样囊性癌细胞系SACC-83中,可见CD44v6表达率为42.47±7.37%,CD133表达率为56.56±10.27%,ABCG2表达率为2.94±1.23%。结论 1.人泪腺腺样囊性癌及人涎腺腺样囊性癌细胞系SACC-83中均有VM结构表达 2.人泪腺腺样囊性癌的病理分型对VM的表达并无直接影响,而CD44v6则与其分型有关； 3.人泪腺腺样囊性癌中VM与CD44v6的表达有关,提示不仅VM与人泪腺腺样囊性癌的转移复发有关,而且CD44v6可能影响VM的生成。 4.人泪腺腺样囊性癌及人涎腺腺样囊性癌细胞系SACC-83中可不同程度的表达肿瘤干细胞相关标志蛋白CD44v6、CD133、ABCG2,提示人泪腺腺样囊性癌中可能存在肿瘤干细胞；
[Abstract]:objective
1., we searched and confirmed the vasculogenic mimicry structure in human adenoid cystadenocarcinoma specimens, and identified its relationship with pathological typing and its significance.
2., we observed the expression of tumor stem cell marker associated protein CD44v6, CD133 and ABCG2 markers in histologic specimens of human adenoid cystadenocarcinoma, analyzed and compared the correlation between vasculogenic mimicry and their correlation, which provided new explanations for the local recurrence of the adenocarcinoma.
3. from the cellular level again were observed, clear angiogenesis exists in structural mimicry of human salivary adenoid cystic carcinoma cell line SACC-83, and to observe the tumor stem cell markers CD44v6, CD133, ABCG2 expression in human salivary adenoid cystic carcinoma cell line SACC-83. Lay the foundation for further tumor mechanism the extraction and analysis of VM generation of stem cells.
Method
1. this experiment collected in Second Hospital Affiliated to Tianjin Medical University in January 1999 33 cases of adenoid cystadenocarcinoma tissues were resected with the experimental conditions of -2009 in March. All specimens were stained by HE after re observation confirmed pathological diagnosis and classification. By CD31/PAS staining under the light microscope observation sections in human lacrimal adenoid cystic carcinoma tissue samples to identify the structure of VM. At the same time the application of two step immunohistochemistry in 33 specimens of CD44v6, expression of CD133 and ABCG2.
2., according to the observation of VM structure in adenoid cystadenocarcinoma of lacrimal gland, we applied statistical methods to compare their expression in different pathological types, and whether there was difference in the expression of CD44v6, CD133 and ABCG2 between VM expression group and non expression group.
3. by human salivary adenoid cystic carcinoma cell line SACC-83 by Li Shenglin the Central Laboratory of Stomatology Hospital of Beijing Medical University) present two-dimensional culture and three-dimensional culture, observation of vasculogenic mimicry and tumor stem cell marker protein CD44v6, CD133, ABCG2 expression in SACC-83 cell line.
Result
1.33 cases were observed by HE staining, and immunohistochemistry results confirmed the diagnosis of lacrimal gland adenoid cystic carcinoma, including 13 cases of type (39.39%), 20 cases of adenoid tubular type (60.61%). By CD31/PAS double staining method to observe the structure of VM, found in 10 samples, the rate was 30.3%., 6 cases of solid type (46.2%), 4 cases of adenoid tubular type (20%), the difference between the two groups was not statistically significant (p=0.346).
2. VM were observed by adenoid cystic carcinoma cells surrounding the formation of tubular, slit shaped structure, the space structure visible red blood cells, red blood cells and the integrity of the structure, not around hemorrhage exist. We also observed the tumor cells cytoplasm of a large number of endocrine PAS positive substance and CD31 positive expression in the VM material. Group and non expression group CD44v6 were 1.57% and 2.51%, two showed a significant difference (p=0.023).
Methods 3.33 cases of tumors were observed by immunohistochemistry of stem cell related markers CD44v6, CD133, ABCG2 expression were CD44v6 positive expression rate was 54.5% (18/33). The positive rate of entity type 76.9% (10/13), the positive rate of adenoid tubular type 40% (8/20), the two groups was statistically significant the difference (p=0.037) positive expression rate of.CD133 was 57.6% (19/33), the entity type and adenoid tubular type were 76.92% (10/13), 45% (9/20), the difference was not statistically significant (p=0.070); protein expression localized in cell membrane and cytoplasm, pale yellow to brown, some of the cases at the same time the expression in the cytoplasm and nucleus of.ABCG2 positive expression rate was 21.2% (7/33), the entity type and adenoid tubular type were 30.8% (4/13), 15% (3/20), the difference was not statistically significant (p=0.297); the positive expression of ABCG2 cells with vascular distribution along the tendency.
4. in the two-dimensional culture of salivary adenoid cystic carcinoma cell line SACC-83 cells were visible, ring, mesh growth, a cavity shaped, three-dimensional culture in tubular growth. In three-dimensional culture showed positive expression of VM. Cancer stem cell related protein in human salivary adenoid cystic carcinoma cell line SACC-83, visible expression of CD44v6 the rate is 42.47 + 7.37%, the expression rate of CD133 was 56.56 + 10.27%, the expression rate of ABCG2 was 2.94 + 1.23%.
conclusion
The expression of VM in 1. human lacrimal gland adenoid cystic carcinoma and human salivary gland adenoid cystic carcinoma cell line SACC-83
The pathological classification of 2. human lacrimal gland adenoid cystic carcinoma has no direct influence on the expression of VM, while CD44v6 is related to its typing.
The expression of VM is correlated with the expression of CD44v6 in 3. cases of adenoid cystic carcinoma of lacrimal gland, suggesting that VM is not only related to metastasis and recurrence of human lacrimal gland adenoid cystic carcinoma, but also CD44v6 may affect the generation of VM.
4. human lacrimal gland adenoid cystic carcinoma and human salivary adenoid cystic carcinoma cell line SACC-83 can express CD44v6, CD133 and ABCG2 of tumor stem cells related to different degrees, suggesting that there may be cancer stem cells in human lacrimal adenoid cystic carcinoma.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.7

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