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Development of APTES-Decorated HepG2 Cancer Stem Cell Membrane Chromatography for Screening Active Components from Salvia miltiorrhiza
Analytical Chemistry ( IF 6.7 ) Pub Date : 2016-11-30 00:00:00 , DOI: 10.1021/acs.analchem.6b02709 Xuan Ding 1 , Yan Cao 1 , Yongfang Yuan 2 , Zhirong Gong 1 , Yue Liu 1 , Liang Zhao 3 , Lei Lv 3 , Guoqing Zhang 3 , Dongyao Wang 1 , Dan Jia 1 , Zhenyu Zhu 1 , Zhanying Hong 1 , Xiaofei Chen 1 , Yifeng Chai 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2016-11-30 00:00:00 , DOI: 10.1021/acs.analchem.6b02709 Xuan Ding 1 , Yan Cao 1 , Yongfang Yuan 2 , Zhirong Gong 1 , Yue Liu 1 , Liang Zhao 3 , Lei Lv 3 , Guoqing Zhang 3 , Dongyao Wang 1 , Dan Jia 1 , Zhenyu Zhu 1 , Zhanying Hong 1 , Xiaofei Chen 1 , Yifeng Chai 1
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Cell membrane chromatography (CMC) is an ideal method for screening potential active components acting on target cell membranes from a complex system, such as herbal medicines. But due to the decay and falling-off of membranes, the CMC column suffers from short life span and low reproducibility. This has greatly limited the application of this model, especially when the cell materials are hard to obtain. To solve this problem, a novel type of (3-aminopropyl)triethoxysilane (APTES)-decorated silica gel was employed. The silica gel was decorated with aldehydes with the help of APTES, which react with the amino groups on cell membranes to form a covalent bond. In this way, cell membranes were immobilized on the surface of silica gel, so it is not easy for membranes to fall off. According to our investigation, the column life of the APTES-decorated group was prolonged to more than 12 days, while the control group showed a sharp decline in column efficiency in the first 3 days. To verify this model, a novel APTES-decorated HepG2 cancer stem cell membrane chromatography (CSCMC) was established and applied in a comprehensive two-dimensional chromatographic system to screen potential active components in Salvia miltiorrhiza. As a result, tanshinone IIA, cryptotanshinone, and dihydrotanshinone I were retained on this model and proved to be effective on HepG2 cancer stem cells by the following cell proliferation and apoptosis assay, with IC50 of 10.30 μM, 17.85 μM, and 2.53 μM, respectively. This improvement of CMC can significantly prolong its column life span and broaden the range of its application, which is very suitable for making invaluable or hard-to-obtain cell materials, such as stem cells, for specific drug screening.
中文翻译：
APTES装饰的HepG2癌症干细胞膜色谱的发展，用于从丹参中筛选活性成分
细胞膜色谱（CMC）是从复杂系统（例如草药）中筛选作用于靶细胞膜的潜在活性成分的理想方法。但是，由于膜的衰减和脱落，CMC色谱柱使用寿命短且重现性差。这极大地限制了该模型的应用，尤其是当难以获得细胞材料时。为了解决该问题，使用了新型的（3-氨基丙基）三乙氧基硅烷（APTES）修饰的硅胶。硅胶在APTES的帮助下用醛修饰，该醛与细胞膜上的氨基反应形成共价键。这样，细胞膜被固定在硅胶表面上，因此膜不容易脱落。根据我们的调查，APTES装饰组的色谱柱寿命延长到超过12天，而对照组在前3天的色谱柱效率急剧下降。为了验证该模型，建立了一种新型的APTES装饰的HepG2癌症干细胞膜色谱法（CSCMC），并在全面的二维色谱系统中进行了应用，以筛选潜在的活性成分。丹参。结果，丹参酮IIA，隐丹参酮和二氢丹参酮I保留在该模型上，并通过以下细胞增殖和凋亡试验证明对HepG2癌症干细胞有效，IC 50为10.30μM，17.85μM和2.53μM，分别。CMC的这一改进可以显着延长其色谱柱寿命，并拓宽了其应用范围，这非常适合于制备用于特定药物筛选的宝贵或难以获得的细胞材料（例如干细胞）。
更新日期：2016-11-30
中文翻译：
APTES装饰的HepG2癌症干细胞膜色谱的发展，用于从丹参中筛选活性成分
细胞膜色谱（CMC）是从复杂系统（例如草药）中筛选作用于靶细胞膜的潜在活性成分的理想方法。但是，由于膜的衰减和脱落，CMC色谱柱使用寿命短且重现性差。这极大地限制了该模型的应用，尤其是当难以获得细胞材料时。为了解决该问题，使用了新型的（3-氨基丙基）三乙氧基硅烷（APTES）修饰的硅胶。硅胶在APTES的帮助下用醛修饰，该醛与细胞膜上的氨基反应形成共价键。这样，细胞膜被固定在硅胶表面上，因此膜不容易脱落。根据我们的调查，APTES装饰组的色谱柱寿命延长到超过12天，而对照组在前3天的色谱柱效率急剧下降。为了验证该模型，建立了一种新型的APTES装饰的HepG2癌症干细胞膜色谱法（CSCMC），并在全面的二维色谱系统中进行了应用，以筛选潜在的活性成分。丹参。结果，丹参酮IIA，隐丹参酮和二氢丹参酮I保留在该模型上，并通过以下细胞增殖和凋亡试验证明对HepG2癌症干细胞有效，IC 50为10.30μM，17.85μM和2.53μM，分别。CMC的这一改进可以显着延长其色谱柱寿命，并拓宽了其应用范围，这非常适合于制备用于特定药物筛选的宝贵或难以获得的细胞材料（例如干细胞）。
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