【作者】 张爽；

【导师】 林贞花；

【作者基本信息】 延边大学 ， 病理学与病理生理学， 2011， 硕士

【摘要】 目的：应用SNP(单核苷酸多态性)基因芯片和免疫组化等实验方法探讨乳腺癌中转移相关基因差异表达和Ezrin基因蛋白过表达的临床病理学意义。材料：选取140例来自延边肿瘤医院病理科、上海同济大学附属第一妇婴医院病理科的乳腺病变组织存档蜡块标本,其中,乳腺癌87例,乳腺原位癌(DCIS)33例,乳腺癌旁正常组织20例。此外,还选取了新鲜乳腺病变组织标本12例,其中,乳腺癌8例(有淋巴结转移、无淋巴结转移各4例)、乳腺纤维瘤1例、乳腺腺病1例和正常乳腺组织2例,均由延边肿瘤医院病理科提供。方法：应用SNP(单核苷酸多态性)基因芯片技术分析乳腺癌中20种转移相关基因的差异表达(拷贝数),同时应用免疫组化EnVision法检测Ezrin蛋白在乳腺癌、DCIS和乳腺癌旁正常组织中的表达水平,并分析其蛋白检测的临床病理学意义。结果：SNP基因芯片检测结果表明,与无淋巴结转移的乳腺癌相比,在淋巴结转移阳性的乳腺癌中Ezrin、BRCA1、SDX4、P27、NEDD9、TSC2、TPM3、ST3 FLT4等9种转移相关基因明显扩增,而RORB、EPHB2、HGF、MYCL1、BRCA2、SHH、PTEN、ESR1、TWIST1、NF2、HDNC1等11种转移相关基因则明显缺失。免疫组化检测结果表明：Ezrin蛋白在有淋巴结转移的乳腺癌中表达率明显增高,阳性和强阳性表达率分别达100%(46/46)和89.1%(43/46)；而在无淋巴结转移的乳腺癌中的表达率则明显下降,阳性和强阳性表达率分别仅为73.2%(30/41)和31.7%%(13/41)(P<0.05)。另外,免疫组化实验结果显示,Ezrin蛋白在癌旁正常组织中的阳性表达率较低,仅为10%(2/20),且主要表现为弱阳性,但在乳腺癌和DCIS中的阳性表达率则明显升高,分别达87.4%(76/87)和87.9%(29/33),强阳性表达率分别达64.4%(56/87)和60.6%(20/33)(P<0.01)。实验还发现,Ezrin蛋白在正常乳腺组织中表达阳性时,其阳性信号主要分布在乳腺腺体中靠近内侧腔面的腺上皮细胞浆中；而在乳腺癌中,Ezrin蛋白表现为弥漫性的阳性染色。结论：乳腺癌转移与多种转移相关基因及其蛋白密切相关。其中,Ezrin基因蛋白在乳腺癌转移和演进过程中起重要作用,是乳腺癌预后和早期诊断的重要检测指标,而有望成为乳腺癌治疗的新靶点。更多还原

【Abstract】 Objectives:To investigate the different expression of metastasis related genes and clinicopathological significance of Ezrin protein overexpression in breast cancer.Materials and Methods:Total 140 cases of paraffin-embedded breast lesion blocks, including 87 cases of breast cancers,33 cases of ductal carcinoma in situ (DCIS), and 20 cases of normal breast tissues, were selected from Department of Pathology, Yanbian Tumor Hospital and The First Maternity and Infant health Hospital, Tongji University School of Medicine. Gene differential expression (copy number) was detected by SNP gene chip for analyzing 20 species genes associated with cell metastasis in breast cancer. Additionally, Ezrin gene protein expression level was detected in normal breast tissues, ductal carcinoma in situ (DCIS), and breast cancer including with or without lymph node metastasis by using EnVision method of immunohistochemistry, and the clinicopathological significance of Ezrin protein overexpression in breast cancer was also analyzed.Results:SNP gene chip results showed that the copy number of Ezrin, BRCA, SDX4, P27, NEDD9, TSC2, TPM3, FLT4, and ST3 were up-regulated, while RORB, EPHB2, HGF, MYCL1, BRCA2, SHH, PTEN, ESR1, Twistl, NF2, and HDAC1 were down-regulated in breast cancers with lymph node metastasis compared with non-metastastatic breast cancers. Immunohistochemical staining showed that the positive and strongly positive rates of Ezrin protein were 100%(46/46) and 89.1%(43/46) in breast cancers with lymph node metastasis, respectively, and 73.2%(30/41) and 31.7%(13/41) in non-metastatic breast cancers, respectively. The difference was significant (P<0.05). Additionally, the positive rate of Ezrin protein was only 10%(2/20), and the positive signals were very weak in normal breast tissues adjacent to cancer, however, the positive rates of Ezrin protein were 87.4%(76/87) and 87.9%(29/33), and the strongly positive rate of Ezrin protein were 64.4%(56/87) and 60.6%(20/33) in breast cancer and DCIS tissues, respectively (P< 0.01). The positive signals mainly distributed at the cytoplasm adjacent to the normal gland lumen, however, Ezrin protein in the breast cancer cells showed diffusely positive staining pattern.Conclusions:Breast cancer metastasis was closely related with a variety of genes and proteins expression. In which, Ezrin gene played an important role in the metastasis and progression of breast cancer. The detection of Ezrin protein might be a useful marker for early diagnosis and prognosis of breast cancer. And Ezrin gene might be the new target for breast cancer therapy.更多还原