聂丽菊, 许恒毅, 叶称连, 黄小林, 刘雯婷, 傅芬. 生物素-链霉亲和素介导受体靶向卵巢癌SKOV3细胞量子点体外成像的研究[J]. 中国肿瘤临床, 2014, 41(19): 1254-1258. DOI: 10.3969/j.issn.1000-8179.20140842
引用本文: 聂丽菊, 许恒毅, 叶称连, 黄小林, 刘雯婷, 傅芬. 生物素-链霉亲和素介导受体靶向卵巢癌SKOV3细胞量子点体外成像的研究[J]. 中国肿瘤临床, 2014, 41(19): 1254-1258. DOI: 10.3969/j.issn.1000-8179.20140842
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NIE1 Liju, XU Hengyi, YE Chenlian, HUANG Xiaolin, LIU Wenting, FU Fen. Biotin-streptavidin system-mediated folate receptor-targeted quantum dot in vitro imaging of ovarian cancer SKOV3 cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2014, 41(19): 1254-1258. DOI: 10.3969/j.issn.1000-8179.20140842
Citation: NIE1 Liju, XU Hengyi, YE Chenlian, HUANG Xiaolin, LIU Wenting, FU Fen. Biotin-streptavidin system-mediated folate receptor-targeted quantum dot in vitro imaging of ovarian cancer SKOV3 cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2014, 41(19): 1254-1258. DOI: 10.3969/j.issn.1000-8179.20140842
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生物素-链霉亲和素介导受体靶向卵巢癌SKOV3细胞量子点体外成像的研究

Biotin-streptavidin system-mediated folate receptor-targeted quantum dot in vitro imaging of ovarian cancer SKOV3 cells

    摘要
  • 摘要:
      目的  研制一种生物素-亲和素系统（BAS）介导叶酸受体（FR）靶向量子点（QD）荧光探针，初步验证其靶向性及信号放大效应。
      方法  采用活泼酯法，将链霉亲和素（SA）与QD共价偶联制备QD-SA并对其物理特性进行验证；采用活泼酯法以牛血清蛋白为载体合成生物素化叶酸（FA）。通过生物素化FA与QD-SA结合制备BAS介导叶酸受体（FR）靶向QD探针，比较该探针对卵巢癌SKOV3细胞及FR表达阴性的肺癌A549细胞的识别情况，并验证其靶向特异性；对比该探针在生物素化FA孵育1、4 h时成像的效果；比较该探针与未经BAS介导的QD-FA探针在不同孵育时间对卵巢癌SKOV3细胞成像的差异。
      结果  BAS介导FR靶向QD探针可特异性识别FR表达阳性的卵巢癌SKOV3细胞，且孵育4 h较1 h荧光信号明显增强，同等条件下其产生的荧光强度较未经BAS介导的QD-FA探针明显增强。
      结论  制备了一种特异性好、灵敏度高的BAS介导FR靶向QD探针，该探针在卵巢癌早期诊断方面具有潜在应用价值。

     

    Abstract:
      Objective   To develop a biotin-streptavidin system (BAS)-mediated folate receptor (FR)-targeted quantum dot (QD) fluorescent probe and preliminarily validate the targeting ability and signal amplification effect of the probe.
      Methods   Streptavidin (SA) was covalently coupled with QD through the active ester method; the physical characteristics of the prepared QD-SA were verified. Biotinylated folate was synthesized through the carrier bovine serum albumin using the same method and then reacted with QD-SA to form the special probe. The probe was used to identify SKOV3 cells and FR-negative A549 cells to verify its targeting specificity. QD-SA was used as the contrast. SKOV3 cells were imaged using the BAS-mediated FR-targeted QD probe with a biotinylated folate incubation time of 1 or 4 h. Various reaction times were also tested between the probe and the QD-FA that was formed without BAS mediation.
      Results   The BAS-mediated FR-targeted QD probe specifically recognized FR-positive SKOV3 cells. The probe obtained higher fluorescent intensity after 4 h than after 1 h of biotinylated folate incubation. The BAS-mediated FR-targeted QD probe also had a stronger fluorescent signal than the QD-FA probe.
      Conclusion   The proposed probe presents a great potential in the early diagnosis of ovarian cancer because of its high specificity and sensitivity.

     

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