Development of APTES-Decorated HepG2 Cancer Stem Cell Membrane Chromatography for Screening Active Components from Salvia miltiorrhiza

Abstract

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 IC 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.

Comprehensive two‐dimensional PC‐3 prostate cancer cell membrane chromatography for screening anti‐tumor components from Radix Sophorae flavescentis

“…To characterize and identify active components interacting with the cell membrane stationary phase, several 2D LC methods have been developed for the rapid investigation of complex TCMs, including heart‐cutting 2D CMC coupled with MS and a comprehensive 2D CMC–MS method . The comprehensive 2D LC–MS system is a powerful analytical technique because of its rapid, efficient separation, high resolution, and powerful identification abilities when coupled with MS, and it is suitable for the analysis of complex TCMs .…”

Label‐free Bio‐affinity Mass Spectrometry for Screening and Locating Bioactive Molecules

“…APTES was modified on the surface of silica, which can provide an amino group to covalently link cell membrane fragments with the help of glutaraldehyde. Ding et al (2016) utilized APTES‐decorated silica gel, which reacts with the amino groups on cell membranes to form a covalent bond. The CMC column life was extended from 3 to 12 days.…”

Recent development in mass spectrometry and its hyphenated techniques for the analysis of medicinal plants

“…Nonetheless, the CMC suffers from the short life span and low reproducibility, which is caused by the decay and falling‐off of membranes . Ding et al . developed a novel type of (3‐aminopropyl) triethoxysilane (APTES)‐decorated silica gel, which can react with the amino groups on cell membranes to form a covalent bond for preventing the falling‐off of membranes.…”