乳腺浸润性导管癌患者血清PF4、VEGF蛋白表达的临床意义

发布时间：2018-04-23 03:26

本文选题：乳腺浸润性导管癌 + PF4　；参考：《河北医科大学》2015年硕士论文

【摘要】：目的:乳腺癌是一种血管依赖性实体肿瘤,它的生长、转移与血管新生密切相关,血管新生受到一系列血管生成调节因子的作用,这些因子控制着血管内皮细胞的生长。血小板第4因子(platelet factor-4,PF4)是一种血小板衍生因子,具有抑制血管生成的作用。研究证实PF4通过抗血管生成作用,抑制肿瘤的发生和发展,但其在乳腺癌中的表达国内外少有报道。血管内皮生长因子(vascular endothelial growth factor,VEGF)是一种强有力的,最具特征性的促血管生成因子,在乳腺癌的发生、生长和转移过程中起重要作用。肿瘤组织和外周血中VEGF水平可以作为肿瘤诊断及评估预后的指标。检测外周血中VEGF的水平更方便,更易于临床应用。CD31即血小板内皮粘附因子,可以标记血管内皮细胞,用来检测微血管密度(microverssel density,MVD),能定量反映乳腺癌组织的血管生长情况。本研究通过检测血清中PF4、VEGF浓度及组织中MVD-CD31表达,来了解PF4、VEGF及MVD-CD31在乳腺浸润性导管癌患者中的变化规律,探讨它们之间及与其他各临床病理特征的关系。方法:1研究对象:选取2014年1月—2014年7月在河北医科大学第四医院乳腺中心就诊,经空芯针穿刺,病理证实为乳腺浸润性导管癌患者60例,均为原发性单侧、未接受过化疗、放疗、内分泌治疗和分子靶向治疗,既往无肿瘤病史的患者。术前抽取空腹静脉血2ml,1000r/min离心3分钟,取上层血清置-80℃冰箱中保存待检测。组织标本在离体30分钟内取肿瘤组织、10%中性福尔马林固定,石蜡包埋待检测。同法用30例乳腺良性疾病(乳腺腺病、纤维腺瘤等)患者的血清和乳腺组织作对照。以上样本的采集均通过河北医科大学第四医院伦理委员会审批。2用酶联免疫吸附试验(ELISA)检测血清中PF4、VEGF蛋白表达,采用美国全自动酶标检测仪VERSAmax,30分钟内在波长450nm的酶标仪上读取各孔OD值。根据OD值绘制Log-logit双对数标准曲线,使用logit-log或四参数数据处理模式(采用SOFTmax PRO4.3LS分析软件),由电脑自动计算得出血清PF4,VEGF结果。3组织切片采用免疫组化SP法染色,经抗原热修复,加入一抗,4℃过夜;加入二抗,室温孵育30min;加入三抗;DAB显色;复染,封片。ER、PR的判读参照2010年ASCO/CAP指南。HER-2的判读参照2007年ASCO/CAP指南。MVD-CD31的MVD判读参照Weidner等报道的方法。4统计分析:采用SPSS11.5统计软件,数据经过方差齐性检验均符合正态分布,计量资料采用均数±标准差表示,两组间均数比较用t检验,三组间均数比较用方差分析,两两比较用LSD法。相关性分析使用Person检验。检验水准α=0.05,P0.05为差异有统计学意义。结果:1乳腺浸润性导管癌患者和对照组血清PF4浓度分别为0.354±0.198ng/ml和0.225±0.436ng/ml,二者差异有统计学意义,P=0.006。乳腺浸润性导管癌患者和对照组血清VEGF浓度分别为15.458±15.819pg/ml和7.727±8.307pg/ml,二者差异有统计学意义,P=0.046。乳腺浸润性导管癌患者和对照组MVD-CD31分别为16.58±9.528和10.40±3.050,二者差异有统计学意义,P=0.005。2乳腺浸润性导管癌患者血清PF4、VEGF之间有显著的相关性,为正相关,r=0.693,P=0.000。血清PF4与MVD-CD31表达无相关性,r=0.01,P=0.947。血清VEGF与MVD-CD31表达无相关性,r=-0.075,P=0.605。3有脉管瘤栓患者的血清PF4表达为0.233±0.089ng/ml,低于无脉管瘤栓者0.396±0.209ng/ml,二者有统计学差异,P=0.009。血清PF4与患者月经状况、肿瘤的大小、TNM分期、组织学分级、淋巴结是否转移、ER、PR、HER-2表达均无相关性,差异均无统计学意义,P0.05。4血清VEGF与患者月经状况无相关性、血清VEGF与肿瘤的大小、TNM分期、组织学分级、淋巴结是否转移、有无脉管瘤栓、ER、PR、HER-2表达均无相关性,差异均无统计学意义,P0.05。5 MVD-CD31在直径2cm的肿瘤组织中表达为21.080±9.358,明显高于直径≤2cm的肿瘤中的12.080±7.455,差异有统计学意义,P=0.000。随着肿瘤TNM分期增高组织中MVD-CD31表达也增高(9.863±5.985,15.304±8.646,22.776±8.348),差异有统计学意义,F=2.983,P=0.002。MVD-CD31与患者月经状况、组织学分级、有无脉管瘤栓、淋巴结是否转移、ER、PR、HER-2表达均无相关性,差异均无统计学意义,P0.05。结论:1 PF4、VEGF在乳腺浸润性导管癌患者血清中浓度明显高于对照组,可以成为鉴别乳腺良、恶性疾病的重要指标。2乳腺浸润性导管癌患者血清PF4、VEGF表达之间呈正相关。3血清PF4、VEGF与MVD-CD31表达无相关性。4 MVD-CD31表达可以反应乳腺浸润性导管癌的生物学行为。
[Abstract]:Objective: breast cancer is a vascular dependent solid tumor. Its growth and metastasis are closely related to angiogenesis. Angiogenesis is affected by a series of angiogenic regulators. These factors control the growth of vascular endothelial cells. Platelet fourth factor (platelet factor-4, PF4) is a kind of platelet derived factor, which has the inhibition of blood. Guan Shengcheng's role. Studies have shown that PF4 inhibits the occurrence and development of tumor by anti angiogenesis effect, but its expression in breast cancer is rarely reported. Vascular endothelial growth factor (VEGF) is a powerful, most specific angiogenic factor, in the occurrence of breast cancer. VEGF levels in tumor tissue and peripheral blood can be used as an indicator of tumor diagnosis and evaluation of prognosis. The detection of VEGF levels in peripheral blood is more convenient and more easy to use.CD31 as a platelet endothelial adhesion factor, which can be used to mark vascular endothelial cells and to detect microvascular density (microverssel densit). Y, MVD) can quantitatively reflect the vascular growth of breast cancer tissues. By detecting the serum levels of PF4, VEGF, and MVD-CD31 expression in the tissues, the changes of PF4, VEGF and MVD-CD31 in patients with invasive ductal carcinoma of the breast were investigated, and the relationship between them and other clinicopathological features was explored. Methods: 1 subjects: 2 014 years from January to July 2014 at the center of the breast center of the fourth hospital of Hebei Medical University, 60 cases of invasive ductal carcinoma of the breast were confirmed by hollow needle puncture. All of the 60 cases of breast invasive ductal carcinoma were primary unilateral, without chemotherapy, radiotherapy, endocrine therapy and molecular targeted therapy. Patients who had no history of cancer were treated with 2ml, 1000r/ Min was centrifuged for 3 minutes, and the upper level serum was stored in the refrigerator at -80 C to be tested. The tissue specimens were collected in 30 minutes in vitro, 10% neutral formalin was fixed, and paraffin embedded to be detected. 30 cases of benign breast disease (mammary gland disease, fibroadenoma, etc.) were compared with the breast tissue. The ethics committee of the fourth hospital of North Medical University examined and approved.2 using enzyme linked immunosorbent assay (ELISA) to detect PF4, VEGF protein expression in serum, use American fully automatic enzyme labeling instrument VERSAmax, and read the orifice of each hole in the enzyme labelled instrument with wavelength 450nm within 30 minutes. The Log-logit double logarithmic standard curve was plotted according to the OD value, and logit-log or four parameters were used. Data processing mode (using SOFTmax PRO4.3LS analysis software), the computer automatically calculated bleeding PF4, VEGF results.3 tissue section using immunohistochemical SP dyeing, antigen heat repair, adding one anti, 4 C overnight; adding two anti, room temperature incubating 30min; adding three resistance; DAB color; re dyeing,.ER, PR interpretation reference 2010 ASCO/CAP guide. HER-2's interpretation refer to the method of.4 statistical analysis, such as the MVD interpretation of the 2007 ASCO/CAP guide.MVD-CD31, and Weidner and other reports. Using SPSS11.5 statistical software, the data are all conformed to normal distribution through the homogeneity test of variance, and the measurement data are represented by mean number + standard deviation. The average number ratio between the two groups is compared with t test and the average of the three groups is compared with the variance analysis, 22 compared with the LSD method. Correlation analysis used Person test. Test level alpha =0.05, P0.05 was statistically significant. Results: 1 the serum PF4 concentrations in patients with invasive ductal carcinoma and control group were 0.354 + 0.198ng/ml and 0.225 + 0.436ng/ml respectively. The two differences were statistically significant, P=0.006. breast infiltrating ductal carcinoma patients and control The serum VEGF concentration was 15.458 + 15.819pg/ml and 7.727 + 8.307pg/ml respectively. The difference between the two groups was statistically significant. The P=0.046. breast invasive ductal carcinoma patients and the control group were 16.58 + 9.528 and 10.40 + 3.050 respectively, and the differences were statistically significant. There was a significant phase between the serum PF4 and VEGF in the patients with P=0.005.2 breast infiltrating ductal carcinoma. There was no correlation between the positive correlation, r=0.693, P=0.000. serum PF4 and MVD-CD31 expression, r=0.01, P=0.947. serum VEGF and MVD-CD31 expression was not related, r=-0.075, P=0.605.3 with vascular tumor suppository in patients with PF4 expression of 0.233 + 0.089ng/ml, lower than the non vascular tumor thrombus 0.396 +, two there were statistical differences. The patients' menstrual condition, tumor size, TNM staging, histological grade, lymph node metastasis, ER, PR, HER-2 expression were not related, the difference was not statistically significant, P0.05.4 serum VEGF was not related to the patient's menstrual status, serum VEGF and tumor size, TNM staging, histological grading, lymph node metastasis, vascular tumor thrombus, ER, PR, HER-2. There was no statistical significance in the expression. The expression of P0.05.5 MVD-CD31 was 21.080 + 9.358 in the tumor tissue of the diameter 2cm, which was significantly higher than 12.080 + 7.455 in the tumor with diameter less than 2cm. The difference was statistically significant. P=0.000. was also increased (9.863 + 5.985,15.304 + 8.646,22.776) with the increase of TNM stage in the tumor. The difference was statistically significant. F=2.983, P=0.002.MVD-CD31 and the patient's menstrual status, histological grading, without vascular tumor thrombus, lymph node metastasis, ER, PR, HER-2 expression were not related, the difference was not statistically significant, P0.05. conclusion: 1 PF4, VEGF in the mammary gland infiltrating ductal carcinoma patients' serum concentration was significantly higher than the control group, can .2 is an important indicator of breast benign and malignant diseases. The serum PF4 in patients with invasive ductal carcinoma of the breast, the positive correlation between the expression of VEGF and the expression of.3 serum PF4, VEGF and MVD-CD31 expression without correlation.4 MVD-CD31 expression can reflect the biological behavior of invasive ductal carcinoma of the breast.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R737.9

【参考文献】