张新颖, 尹富强, 刘丽, 高婷, 阮和云, 冠潇, 卢迎新, 黎丹戎. 人淋巴管内皮细胞对卵巢癌淋巴结定向转移细胞分泌蛋白表达的分析[J]. 中国肿瘤临床, 2013, 40(19): 1155-1159. DOI: 10.3969/j.issn.1000-8179.20130624
引用本文: 张新颖, 尹富强, 刘丽, 高婷, 阮和云, 冠潇, 卢迎新, 黎丹戎. 人淋巴管内皮细胞对卵巢癌淋巴结定向转移细胞分泌蛋白表达的分析[J]. 中国肿瘤临床, 2013, 40(19): 1155-1159. DOI: 10.3969/j.issn.1000-8179.20130624
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Xinying ZHANG, Fuqiang YIN, Li LIU, Ting GAO, Heyun RUAN, Xiao GUAN, Yingxin LU, Danrong LI. Analysis of the effect of human lymphatic endothelial cells on proteins secreted by epithelial ovarian cancer cells with lymph node-oriented metastasis[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2013, 40(19): 1155-1159. DOI: 10.3969/j.issn.1000-8179.20130624
Citation: Xinying ZHANG, Fuqiang YIN, Li LIU, Ting GAO, Heyun RUAN, Xiao GUAN, Yingxin LU, Danrong LI. Analysis of the effect of human lymphatic endothelial cells on proteins secreted by epithelial ovarian cancer cells with lymph node-oriented metastasis[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2013, 40(19): 1155-1159. DOI: 10.3969/j.issn.1000-8179.20130624
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人淋巴管内皮细胞对卵巢癌淋巴结定向转移细胞分泌蛋白表达的分析

Analysis of the effect of human lymphatic endothelial cells on proteins secreted by epithelial ovarian cancer cells with lymph node-oriented metastasis

    摘要
  • 摘要:
      目的  初步探讨人淋巴管内皮细胞(HLEC)对卵巢癌淋巴结定向转移细胞(SKOV3-PM4)分泌蛋白的影响。
      方法  将细胞体系分为4组(A组：SKOV3；B组：SKOV3 + HLEC；C组：SKOV3-PM4；D组：SKOV3-PM4 + HLEC)，采用抗体芯片和iTRAQ-2D-LC-MALDI-TOF/TOF/MS方法检测各组细胞培养基中的蛋白，筛选出差异明显的分泌蛋白，对其行生物信息学分析，并采用ELISA方法在人血清标本和细胞培养基中进行验证。
      结果  抗体芯片与质谱分析结果提示，C组与A组相比、D组与C组相比Progranulin(GRN)、VEGFA表达上调，SPARC、IGFBP7表达下调。经过综合生物信息学分析，IGFBP7和VEGFA联系紧密，相互影响。与正常组血清相比，VEGFA在卵巢癌中的表达最高，IGFBP7表达最低，差异有统计学意义(P < 0.05)。
      结论  淋巴结定向转移与淋巴管内皮细胞微环境关系密切，共培养后的差异蛋白涉及到基质细胞蛋白、细胞黏附作用因子等方面。其中VEGFA、GRN的表达上调及SPARC、IGFBP7的表达下调与卵巢癌淋巴结定向高转移密切相关。

     

    Abstract:
      Objective   This study aimed to investigate the effect of human lymphatic endothelial cells (HLECs) on proteins secreted by epithelial ovarian cancer (EOC) cells SKOV3-pm4 with highly directional lymphatic metastasis.
      Methods   The supernatants of the four groups of cultured cells (A, SKOV3; B, SKOV3+HLEC; C, SKOV3-PM4; and D, SKOV3-PM4+HLEC) were collected. The proteins of these cells were detected by antibody arrays and iTRAQ-2D-LC-MALDI-TOF/TOF/MS. The screened significantly differential proteins were further analyzed by bioinformatics and validated in the human serum and cell culture medium by ELISA.
      Results   Progranulin (GRN) and vascular endothelial growth factor A (VEGF-A) were upregulated between groups C and A. In addition, insulin-like growth factor binding protein-7 (IGFBP-7) and secreted protein acid rich in cysteine (SPARC) were downregulated between groups D and C. Comprehensive bioinformatics analysis revealed that IGFBP7 interacted with VEGFA. VEGF exhibited the highest expression in ovarian cancer and IGFBP7 exhibited the lowest expression compared with the serum of the normal control group. Statistically significant differences were observed between the two substances.
      Conclusion   The HLEC microenvironment is closely associated with directional metastasis in lymph nodes with differential proteins, including matricellular proteins and adhesion factors. In particular, the upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 were closely associated with the directional metastasis of EOC cells in lymph nodes.

     

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