【作者】 周隽；

【导师】 吴强； 孟刚；

【作者基本信息】 安徽医科大学 ， 病理学与病理生理学， 2003， 硕士

【摘要】 目的：观察乳腺良、恶性病变中抑癌基因PTEN及其蛋白表达的改变，并观察它与乳腺癌临床病理特征的关系，及其与integrinβ1、MMP-2、MMP-9、TIMP-1及细胞周期调控因子cyclinD1之间表达的相关性，探讨其在乳腺癌浸润转移发生发展中的生物学意义。方法：用免疫组织化学S-P法检测23例癌旁正常乳腺组织、28例乳腺增生组织(其中20例普通型增生，8例非典型增生)、8例导管原位癌、98例浸润性乳腺癌组织PTEN蛋白的表达，检测98例浸润癌组织中integrinβ1、MMP-2、MMP-9、TIMP-1、cyclinβ1蛋白的表达，并进行微血管密度计数。应用聚合酶链式反应(PCR)-聚丙烯酰胺尿素凝胶电泳(PAUGE)方法，选择紧邻PTEN侧翼的二核苷酸重复序列D10S215微卫星多态位点，对98例浸润性乳腺癌标本进行有关PTEN微卫星的杂合性缺失和微卫星不稳定性的研究。结果：1．正常乳腺组织的PTEN多呈弱阳性表达(65.4％)，非典型增生上皮和导管原位癌的PTEN蛋白水平高于正常乳腺腺上皮(P＜0.01)。浸润癌组织中PTEN的失表达率为7.1％，高表达率为49.0％，与正常乳腺组织比较有显著性差异(P＜0.05)，与增生的乳腺组织和导管原位癌组间无显著差异。浸润性乳腺癌中PTEN表达与组织学分级、腋淋巴结转移、TNM分期、微血管密度计数均呈显著负相关。2．98例浸润性乳腺癌标本中D10S215位点上32例(32.7％)PTEN有杂合性缺失，其中5例PTEN表达阴性。随腋淋巴结的受累、TNM分期的增高，PTEN的杂合性缺失发生率增高(P＜0.05)。D10S215位点上未检测到微卫星不稳定。3．PTEN蛋白与cyclinD1、integrinβ1、MMP-2、MMP-9蛋白表达呈显著负相关，与TIMP-1蛋白表达呈显著正相关。4．PTEN表达低下组乳腺癌中cyclinD1蛋白的高表达率与腋淋巴结受累、组织学分级呈正相关，integrinβ1蛋白的表达率与腋窝淋巴结、组织学分级无关，MMP-2、MMP-9的表达率与腋窝淋巴结的受累呈正相关，TIMP-1蛋白表达阳性率与腋窝淋巴结的受累无关。5．PTEN高表达组中cyclinD1蛋白的安徽医科大学硕士学位论文周隽·2003高表达率与腋淋巴结受累、组织学分级无关，iniegrin印、M州田一2、MMp一9蛋白的表达率与腋窝淋巴结的受累呈正相关(P<0.01)，TIMP一1与腋窝淋巴结的受累呈负相关。结论:1.PTEN表达的改变是乳腺癌发生的早期事件，PTEN与细胞周期调控因子cyclinDI共同参与乳腺癌的发生。PTEN的表达水平低下可作为乳腺癌分化和转移的一个潜在的生物学标志。2.PTEN基因LOH是乳腺癌发生晚期阶段的重要改变之一，它在乳腺癌的进展和转移过程中可能发挥重要作用。乳腺癌中DIOSZ15位点上未检测到微卫星不稳定，提示微卫星不稳定可能不是浸润性乳腺癌的主要改变。3.浸润性乳腺癌中，免疫组化检测PTEN蛋白表达能较好的反映其基因DlosZ15位点的杂合性缺失。4.CychnDI对乳腺癌分化和腋淋巴结转移的影响可能部分的受到PTEN的调控。5.MMP一2、MMP一9对乳腺癌淋巴结转移的影响作用独立于 PTEN，但TIMP一1、inieghnpl对乳腺癌浸润转移过程的作用可能在一定程度上受到PTEN的调控。更多还原

【Abstract】 Purpose To investigate the change of PTEN gene and its protein expression in breast cancer, study its correlations with integrin 1, MMP-2, MMP-9, TIMP-1 and cyclinDl, so as to understand its roles in tumor invasion and metastasis and to evaluate its clinicopathological significance. Methods Immunohistochemical staining, S-P method, was used to detect the positivity of PTEN in 23 cases of normal breast tissues, 28 cases of proliferative breast disease including 8 cases of atypical hyperplasia (ADH), 8 cases of ductal carcinoma in situ (DCIS) and 98 cases of breast cancer. Meanwhile, cyclinDl, integrin l, MMP-2, MMP-9, TIMP-1 and microvessel density(MVD) marked with Factor VIII-related antigen were also investigated in 98 cases of breast cancer. PCR-PAUGE was used to detect loss of heterozygosity (LOH) and microsatellite instability (MSI) near PTEN loci in 98 cases of breast cancer. Results 1. PTEN was weakly expressed in normal breast tissues. Atypical hyperplasia and DCIS expressed higher levels of PTEN than normal tissues(P<0.05), but had no remarkable distinction with invasive breast cancer. The expression of PTEN was negatively correlated to histological grading, axillary lymph node metastasis, clinical staging and MVD. 2. The frequency of loss of flanking marker D10S215 around PTEN was 32.7%(32/98). In the cases with PTEN-LOH, 5 of 32 were PTEN-negative. PTEN-LOH was positive correlated to clinical staging, axillary lymph node metastasis. But microsatellite analysis showed that there was no microsatellite instability about D10S215 of PTEN gene existed in all these samples. 3. A negative relationship existed between the expression of PTEN and cyclinDl, integrinpl, MMP-2, MMP-9. And a positive relationship existed between the expression of PTEN and TIMP-1. 4. With weak and no expression of PTEN, there was a positive correlation between the expression of cyclinDl andhistological grading, axillary lymph node metastasis, a positive correlation between the expression of MMP-2, MMP-9 and axillary lymph node metastasis, but no correlations between the expression of integrinβ1, TIMP-1 and axillary lymph node metastasis. 5. With strong expression of PTEN, there was a positive correlation between the expression of integrinβ1, MMP-2, MMP-9 and axillary lymph node metastasis, a negative correlation between TIMP-1 expression and axillary lymph node metastasis, but no correlations between cyclinDl expression and axillary lymph node metastasis. Conclusion 1. The change of PTEN expression is an early event in malignant transformation of breast epithelium. Significant correlation between the expression of PTEN and cyclinDl also suggests that the down regulation of PTEN expression in breast cancer correlating with histological grading, axillary lymph node metastasis, clinical stage and MVD may be considered as a potential indicator to judge the differentiation and metastasis of breast cancer. 2. LOH of PTEN gene may be a relatively late event in breast cancer development, which may play important roles on invasion and metastasis in breast cancer. And no microsatellite instability about D10S215 of PTEN gene suggests that it might not be the main change in breast cancer. 3. Immunohistochemistry is useful for the detection of LOH of PTEN gene on D10S215 in breast cancer. 4. The effect of cyclinDl on axillary lymph node metastasis of breast cancer may partly be under the regulation of PTEN. 5. There is a PTEN-independent regulation pathway for MMP-2 and MMP-9 on metastasis and invasion in breast cancer. However, the effect of TIMP-1 and integrinβl may be under the regulation of PTEN.更多还原