CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing, 100049, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Sci China Life Sci. 2018 Apr;61(4):492-500. doi: 10.1007/s11427-017-9261-9. Epub 2018 Mar 14.
Membrane type 1 matrix metalloproteinase (MT1-MMP) is expressed in most tumors and is believed to play a key role in the development, invasion and metastasis of tumors. There is an urgent need to develop a simple method to detect the MT1-MMP expression level on cells. In this current study, we demonstrated a red emission Au cluster probes with the specific targeting of MT1-MMP on human dopaminergic neuroblastoma (SH-SY5Y) cells. More importantly, utilizing the intrinsic enzyme-like activity of the Au cluster probes, the expression level of MT1-MMP on the SH-SY5Y cells could be assessed by the naked eye without cell lysis and protein extraction process. Furthermore, SH-SY5Y, human breast cancer (MCF-7), and human bronchial epithelial (16-HBE) cell lines with different MT1-MMP expression level could be distinguished using the Au cluster probes by the naked eye. Meanwhile, fluorescence intensity and Au count determined by inductively coupled plasma mass spectrometry (ICP-MS) were used to verify the feasibility of this simple analytical method. Our proposed method is rapid, convenient, and accurate and could be assessed by the naked eye. This visual assessment method of tumor-associated proteins has immense implications in sorting tumor cells.
膜型 1 基质金属蛋白酶（MT1-MMP）在大多数肿瘤中表达，被认为在肿瘤的发生、浸润和转移中起关键作用。因此，迫切需要开发一种简单的方法来检测细胞上的 MT1-MMP 表达水平。在本研究中，我们展示了一种红色发射的 Au 团簇探针，该探针对人多巴胺能神经母细胞瘤（SH-SY5Y）细胞具有特定的 MT1-MMP 靶向性。更重要的是，利用 Au 团簇探针的内在酶样活性，无需细胞裂解和蛋白质提取过程，即可通过肉眼评估 SH-SY5Y 细胞上 MT1-MMP 的表达水平。此外，还可以通过肉眼利用 Au 团簇探针区分具有不同 MT1-MMP 表达水平的 SH-SY5Y、人乳腺癌（MCF-7）和人支气管上皮（16-HBE）细胞系。同时，通过电感耦合等离子体质谱法（ICP-MS）测定的荧光强度和 Au 计数验证了该简单分析方法的可行性。我们提出的方法快速、方便、准确，可通过肉眼评估。这种肿瘤相关蛋白的可视化评估方法在肿瘤细胞分选方面具有重要意义。
