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乳腺浸潤性導(dǎo)管癌患者血清PF4、VEGF蛋白表達(dá)的臨床意義

發(fā)布時間:2018-04-23 03:26

  本文選題:乳腺浸潤性導(dǎo)管癌 + PF4 ; 參考:《河北醫(yī)科大學(xué)》2015年碩士論文


【摘要】:目的:乳腺癌是一種血管依賴性實(shí)體腫瘤,它的生長、轉(zhuǎn)移與血管新生密切相關(guān),血管新生受到一系列血管生成調(diào)節(jié)因子的作用,這些因子控制著血管內(nèi)皮細(xì)胞的生長。血小板第4因子(platelet factor-4,PF4)是一種血小板衍生因子,具有抑制血管生成的作用。研究證實(shí)PF4通過抗血管生成作用,抑制腫瘤的發(fā)生和發(fā)展,但其在乳腺癌中的表達(dá)國內(nèi)外少有報(bào)道。血管內(nèi)皮生長因子(vascular endothelial growth factor,VEGF)是一種強(qiáng)有力的,最具特征性的促血管生成因子,在乳腺癌的發(fā)生、生長和轉(zhuǎn)移過程中起重要作用。腫瘤組織和外周血中VEGF水平可以作為腫瘤診斷及評估預(yù)后的指標(biāo)。檢測外周血中VEGF的水平更方便,更易于臨床應(yīng)用。CD31即血小板內(nèi)皮粘附因子,可以標(biāo)記血管內(nèi)皮細(xì)胞,用來檢測微血管密度(microverssel density,MVD),能定量反映乳腺癌組織的血管生長情況。本研究通過檢測血清中PF4、VEGF濃度及組織中MVD-CD31表達(dá),來了解PF4、VEGF及MVD-CD31在乳腺浸潤性導(dǎo)管癌患者中的變化規(guī)律,探討它們之間及與其他各臨床病理特征的關(guān)系。方法:1研究對象:選取2014年1月—2014年7月在河北醫(yī)科大學(xué)第四醫(yī)院乳腺中心就診,經(jīng)空芯針穿刺,病理證實(shí)為乳腺浸潤性導(dǎo)管癌患者60例,均為原發(fā)性單側(cè)、未接受過化療、放療、內(nèi)分泌治療和分子靶向治療,既往無腫瘤病史的患者。術(shù)前抽取空腹靜脈血2ml,1000r/min離心3分鐘,取上層血清置-80℃冰箱中保存待檢測。組織標(biāo)本在離體30分鐘內(nèi)取腫瘤組織、10%中性福爾馬林固定,石蠟包埋待檢測。同法用30例乳腺良性疾病(乳腺腺病、纖維腺瘤等)患者的血清和乳腺組織作對照。以上樣本的采集均通過河北醫(yī)科大學(xué)第四醫(yī)院倫理委員會審批。2用酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測血清中PF4、VEGF蛋白表達(dá),采用美國全自動酶標(biāo)檢測儀VERSAmax,30分鐘內(nèi)在波長450nm的酶標(biāo)儀上讀取各孔OD值。根據(jù)OD值繪制Log-logit雙對數(shù)標(biāo)準(zhǔn)曲線,使用logit-log或四參數(shù)數(shù)據(jù)處理模式(采用SOFTmax PRO4.3LS分析軟件),由電腦自動計(jì)算得出血清PF4,VEGF結(jié)果。3組織切片采用免疫組化SP法染色,經(jīng)抗原熱修復(fù),加入一抗,4℃過夜;加入二抗,室溫孵育30min;加入三抗;DAB顯色;復(fù)染,封片。ER、PR的判讀參照2010年ASCO/CAP指南。HER-2的判讀參照2007年ASCO/CAP指南。MVD-CD31的MVD判讀參照Weidner等報(bào)道的方法。4統(tǒng)計(jì)分析:采用SPSS11.5統(tǒng)計(jì)軟件,數(shù)據(jù)經(jīng)過方差齊性檢驗(yàn)均符合正態(tài)分布,計(jì)量資料采用均數(shù)±標(biāo)準(zhǔn)差表示,兩組間均數(shù)比較用t檢驗(yàn),三組間均數(shù)比較用方差分析,兩兩比較用LSD法。相關(guān)性分析使用Person檢驗(yàn)。檢驗(yàn)水準(zhǔn)α=0.05,P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:1乳腺浸潤性導(dǎo)管癌患者和對照組血清PF4濃度分別為0.354±0.198ng/ml和0.225±0.436ng/ml,二者差異有統(tǒng)計(jì)學(xué)意義,P=0.006。乳腺浸潤性導(dǎo)管癌患者和對照組血清VEGF濃度分別為15.458±15.819pg/ml和7.727±8.307pg/ml,二者差異有統(tǒng)計(jì)學(xué)意義,P=0.046。乳腺浸潤性導(dǎo)管癌患者和對照組MVD-CD31分別為16.58±9.528和10.40±3.050,二者差異有統(tǒng)計(jì)學(xué)意義,P=0.005。2乳腺浸潤性導(dǎo)管癌患者血清PF4、VEGF之間有顯著的相關(guān)性,為正相關(guān),r=0.693,P=0.000。血清PF4與MVD-CD31表達(dá)無相關(guān)性,r=0.01,P=0.947。血清VEGF與MVD-CD31表達(dá)無相關(guān)性,r=-0.075,P=0.605。3有脈管瘤栓患者的血清PF4表達(dá)為0.233±0.089ng/ml,低于無脈管瘤栓者0.396±0.209ng/ml,二者有統(tǒng)計(jì)學(xué)差異,P=0.009。血清PF4與患者月經(jīng)狀況、腫瘤的大小、TNM分期、組織學(xué)分級、淋巴結(jié)是否轉(zhuǎn)移、ER、PR、HER-2表達(dá)均無相關(guān)性,差異均無統(tǒng)計(jì)學(xué)意義,P0.05。4血清VEGF與患者月經(jīng)狀況無相關(guān)性、血清VEGF與腫瘤的大小、TNM分期、組織學(xué)分級、淋巴結(jié)是否轉(zhuǎn)移、有無脈管瘤栓、ER、PR、HER-2表達(dá)均無相關(guān)性,差異均無統(tǒng)計(jì)學(xué)意義,P0.05。5 MVD-CD31在直徑2cm的腫瘤組織中表達(dá)為21.080±9.358,明顯高于直徑≤2cm的腫瘤中的12.080±7.455,差異有統(tǒng)計(jì)學(xué)意義,P=0.000。隨著腫瘤TNM分期增高組織中MVD-CD31表達(dá)也增高(9.863±5.985,15.304±8.646,22.776±8.348),差異有統(tǒng)計(jì)學(xué)意義,F=2.983,P=0.002。MVD-CD31與患者月經(jīng)狀況、組織學(xué)分級、有無脈管瘤栓、淋巴結(jié)是否轉(zhuǎn)移、ER、PR、HER-2表達(dá)均無相關(guān)性,差異均無統(tǒng)計(jì)學(xué)意義,P0.05。結(jié)論:1 PF4、VEGF在乳腺浸潤性導(dǎo)管癌患者血清中濃度明顯高于對照組,可以成為鑒別乳腺良、惡性疾病的重要指標(biāo)。2乳腺浸潤性導(dǎo)管癌患者血清PF4、VEGF表達(dá)之間呈正相關(guān)。3血清PF4、VEGF與MVD-CD31表達(dá)無相關(guān)性。4 MVD-CD31表達(dá)可以反應(yīng)乳腺浸潤性導(dǎo)管癌的生物學(xué)行為。
[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.

【學(xué)位授予單位】:河北醫(yī)科大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2015
【分類號】:R737.9

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 吉曉濱;血小板與腫瘤細(xì)胞的相互作用[J];國外醫(yī)學(xué).耳鼻咽喉科學(xué)分冊;1997年01期

2 張永勝;徐龍江;李峰;馮一中;;胰腺癌組織中EGFR和CD31的表達(dá)及其臨床意義[J];臨床與實(shí)驗(yàn)病理學(xué)雜志;2011年10期

3 鄭玲,王曉毓,張鴻彬,伍建容,谷焰;肺癌患者血小板粘附分子CD31和CD62的檢測[J];中國腫瘤臨床;2003年08期

4 崔曉楠;侯力;;組織微陣列方法檢測CD31、CD105、v-WF、PCNA在肝癌組織中的表達(dá)[J];中國腫瘤;2007年10期

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