标题：MCP-1在乳腺恶性肿瘤细胞增殖与转移中的作用

      作者：吕明丽，周赟，周雷平，牛建梅    (上海交通大学医学院附属国际和平妇幼保健院超声科，上海 200030)

      卷次： 2017年28卷6期

      【摘要】 目的 探讨人单核细胞趋化因子-1 (MCP-1)在乳腺恶性肿瘤细胞增殖转移中的作用。方法 构建MCP-1过表达的稳转人乳腺癌细胞MCF-7细胞和空载MCF-7细胞，分别标记为MCP-1过表达MCF-7细胞(MCP-MCF-7)、空载MCF-7细胞(EV-MCF-7)。通过CCK8细胞增殖实验、单克隆形成实验、划痕实验和细胞凋亡检测，观察MCP-1过表达对细胞增殖转移及凋亡的影响。通过 real-time PCR法检测MCP-1可能的下游基因。结果 成功构建了MCP-1过表达MCF-7乳腺癌细胞株及对照细胞株。CCK8实验显示培养第 5天起MCP-MCF-7细胞株在450 nm处的吸光度明显高于EV-MCF-7组，且差异有统计学意义(P<0.05)。细胞集落形成实验提示MCP-MCF-7细胞株所形成的集落数量明显多于EV-MCF-7细胞，差异有统计学意义(P<0.05)，且前者单个集落的体积亦大于后者。划痕实验中12 h后MCP-MCF-7细胞的迁移能力开始增强，到48 h时划痕更加愈合明显。 虽然转染MCP-1后细胞有效凋亡百分比(0.57%)低于空载细胞(1.10%)，但差异无统计学意义(P>0.05)，即MCP-1对MCF-7乳腺癌细胞凋亡无影响。在对MCP-1下游可能基因的分析结果提示MCP-1可以上调 p65、MMP2、STAT2基因表达，下调RASSF1基因表达。结论 MCP-1具有直接促进乳腺恶性肿瘤细胞的增殖和转移能力，且此种作用可能与p65、MMP2、STAT2、RASSF1有关。
      【关键词】 人单核细胞趋化蛋白-1；乳腺肿瘤细胞；增殖；转移
      【中图分类号】 R737.9 【文献标识码】 A 【文章编号】 1003—6350(2017)06—0861—06

Function of MCP-1 in malignant breast carcinoma cells proliferation and metastasis.
LV Ming-li, ZHOU Yun,ZHOU Lei-ping, NIU Jian-mei. Department of Ultrasonography, International Peace Maternity & Child Health Hospital,School of Medicine, Shanghai Jiaotong University, Shanghai 200030, CHINA
【Abstract】 Objective To analyze the function of monocyte chemotactic protein-1 (MCP-1) in promoting pro-liferation and metastasis of malignant breast cancer cells. Methods MCF-7 cells with MCP-1 over-expression andno-load MCF-7 cells (control) were constructed, which were labeled as MCP-MCF-7 and EV-MCF-7 respectively. TheCCK8 cells proliferation assay, colony formation assay, scratch assay and cell apoptosis assay were employed to mea-sure the effect of MCP-1 over-expression on MCF-7 cells proliferation, metastasis and apoptosis. Real-time PCR wasused to identify candidate downstream genes. Results The MCP-1 over-expressed MC-7 cells and control cell linewere successfully constructed. CCK8 assay showed that the absorbance of MCP-MCF-7 at 450 nm was higher than thatof EV-MCF-7 from day 5, and the statistical significance was significant (P<0.05). Colony formation assay illustratedthat colony number was significantly higher in MCP-MCF-7 cells than the EV-MCF-7 cells (P<0.05), and colony vol-ume was larger in MCP-MCF-7 cells. Scratch assay demonstrated that the migratory ability of MCP-MCF-7 cell in-creased at 12 hours, and wound healing became more evident at 48 hours. The percentage of early apoptoticMCP-MCF-7 cells (0.57%) was lower than that in EV-MCF-7 cells (1.10%), with no statistically significant difference(P>0.05), which suggested no effect on MCP-1 induced apoptosis in MCF-7 cells. Real-time PCR analysis of theMCP-1 downstream-regulated gene indicated that p65, MMP2 and STAT2 gene were up-regulated, while RASSF1 genewas down-regulated. Conclusion MCP-1 directly promotes breast carcinoma cell proliferation and metastasis. This ef-fect may relate to candidate genes p65, MMP2, STAT2 and RASSF1.
      【Key words】 MCP-1; Breast carcinoma cells; Proliferation; Metastasis·论 著·doi:10.3969/j.issn.1003-6350.2017.06.001基金项目：国家自然科学基金(编号：81402386)；中国福利会国际和平妇幼保健院优秀青年培育计划