郭志峰

[摘要] 目的 探討血清中期因子（MK）、血小板因子4（PF4）及黏蛋白1（MUC1）的含量检测在乳腺癌患者病情评估中的应用价值。 方法 选取2012年10月～2016年2月内蒙古自治区赤峰市医院（以下简称“我院”）肿瘤内科收治的180例乳腺肿瘤患者作为观察对象，依据组织病理分型将其分为乳腺良性肿瘤（90例）、乳腺癌（90例）。其中90例乳腺癌患者依据TNM分期分为Ⅰ/Ⅱ期（40例）、Ⅲ/Ⅳ期（50例）；根据分化程度分为高/中分化（50例）、低/未分化（40例）；根据淋巴结转移情况分为淋巴结未转移（50例）、淋巴结转移（40例）。将同期在我院接受体检的120名健康女性作为对照组。收集血清，比较各组之间血清MK、PF4以及MUC1的含量。实时荧光定量PCR（qPCR）检测乳腺癌细胞内MK、PF4以及MUC1过表达对基质金属蛋白酶（MMP）2和MMP9表达的影响，酶联免疫吸附测定（ELISA）检测过表达MK、PF4以及MUC1对细胞上清中血管内皮生长因子（VEGF）A、VEGFB和VEGFC含量的影响。 结果 乳腺良性肿瘤患者MK和PF4含量明显高于对照组（P < 0.05），乳腺癌患者血清中MK、PF4及MUC1的含量明显高于乳腺良性肿瘤患者（P < 0.05）；Ⅲ/Ⅳ期、低/未分化、淋巴结转移患者血清中MK、PF4以及MUC1含量明显高于Ⅰ/Ⅱ期、高/中分化、淋巴结未转移的患者（P < 0.05）；过表达MK、PF4以及MUC1组细胞内MMP2和MMP9的mRNA含量明显高于空载体对照组（P < 0.05）；过表达MK、PF4以及MUC1组细胞培养上清中VEGFA、VEGFB、VEGFC的含量明显高于空载体对照组（P < 0.05）。 结论 乳腺癌患者血清中MK、PF4以及MUC1异常升高；MK、MUC1过表达可能通过调节MMP2、MMP9以及VEGFA、VEGFB、VEGFC的表达影响血管新生和乳腺癌的侵袭。MK、PF4和MUC1可作为新的判定乳腺癌生物学行为的血清标志物。

[关键词] 乳腺癌；中期因子；血小板因子；黏蛋白；基质金属蛋白酶；血管内皮生长因子

[中图分类号] R737.9 [文献标识码] A [文章编号] 1673-7210（2017）04（b）-0099-04

[Abstract] Objective To investigate the application value of content determination of serum midkine （MK）， platelet factor 4 （PF4） and mucoprotein 1 （MUC1） in the disease assessment of patients with breast cancer. Methods One hundred and eighty patients with breast cancer admitted to Department of Medical Oncology， Chifeng Hospital （“our hospital” for short） from October 2012 to February 2016 were selected as research objects， and they were divided into benign breast tumor （90 cases） and breast cancer （90 cases）. The 90 cases with breast cancer were divided into Ⅰ/Ⅱ stage （40 cases） and Ⅲ/Ⅳ stage （50 cases） according to the TNM staging； they were divided into high/medium differentiation （50 cases） and low/undifferentiated （40 cases） dependent on differentiation grade； they were divided into without lymphatic metastasis （50 cases）， lymphatic metastasis （40 cases） according the conditions of lymphatic metastasis. Meanwhile， 120 cases of healthy women taken physical examination in our hospital at the same time were recruited as control group. The serum was collected， and the contents of MK， PF4 and MUC1 in all groups were compared. The effects of over-expression of MK， PF4 and MUC1 in the breast cancer cells for the expression of matrix metalloproteinase （MMP）2 and MMP9 were detected by real-time quantitative PCR （qPCR）. The effects of over-expression of MK， PF4 and MUC1 for the contents of vascular endothelial growth factor （VEGF）A， VEGFB and VEGFC in cellular supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. Results The contents of serum MK and PF4 in patients with benign breast tumor were significantly higher than those of control group （P < 0.05）. While， the contents of serum MK， PF4 and MUC1 in patients with breast cancer were significantly higher than those of patients with benign breast tumor （P < 0.05）. The contents of serum MK， PF4 and MUC1 in patients with Ⅲ/Ⅳ stage， low/undifferentiated and lymphatic metastasi were significantly higher than those of patients with Ⅰ/Ⅱ stage， high/medium differentiation and without lymphatic metastasis （P < 0.05）. The contents of mRNA of MMP2 and MMP9 in the cells with over-expression of MK， PF4 and MUC1 were significantly higher than that of empty gsensor control group （P < 0.05）. Moreover， the contents of VEGFA， VEGFB and VEGFC in supernatant of cells with over-expression of MK， PF4 and MUC1 were significantly higher than that of empty gsensor control group （P < 0.05）. Conclusion The contents of serum MK， PF4 and MUC1 in patients with breast cancer were absolutely increased. Over-expression of MK and PF4 affects angiogenesis and the invasion of breast cancer may be through regulating the expression of MMP2， MMP9 and VEGFA， VEGFB， VEGFC. MK， PF4 and MUC1 can be as the new serum markers to judge the biological behavior of breast cancer.

[Key words] Breast cancer； Midkine； Platelet factor； Mucoprotein； Matrix metalloproteinase； Vascular endothelial growth factor

乳腺癌为女性最常见的恶性肿瘤[1]，其诊疗的关键是早期诊断，目前，常通过检测肿瘤组织中的相关分子进行病理分期和肿瘤恶性程度的判定[2]。肿瘤组织的获取需要穿刺活检，属于有创检查的范畴，不适合病情的长期监测和评估[3]；并且，一般影像学能查出时肿瘤灶已较大，穿刺活检在发现早期肿瘤并进行诊断方面存在弊端，迫切需要新的临床诊断标志物。中期因子（MK）在不同肿瘤中呈明显高表达，且与肿瘤发生、发展、血管形成及转移密切相关，严重影响预后[4]。……