Abstract
BACKGROUND: Recently, the identification of cell apoptosis induced by natural products has become research hotspot and frontier in the biopharmaceutical and food industries under the umbrella of global green development worldwide. Traditionally, cell apoptosis is identified using morphological, biochemical, and cell cycle experiments, which is time consuming, and experimental materials are not from the same group, and it is very hard to ensure the identity and veracity of results of former and latter experiments. AIM: To establish a selective, instant, and practical protocol to identify cell apoptosis induced by natural products. METHODS: A one transient cell processing procedure (OTCPP) was used to detect human colorectal cancer LoVo cell apoptosis after treatment with Pinus massoniana bark extract (PMBE) at the morphological, biochemical, and cell cycle levels. The methods used included treatment with DNA gel electrophoresis, fluorescence microscopy, and flow cytometry. RESULTS: In PMBE-treated LoVo cells, we observed a DNA ladder on gel electrophoresis and fluorescence microscopy revealed "nuclear shrinkage, chromatin condensation or fragmentation". In addition, flow cytometry showed an "obvious apoptosis curve". Thus OTCPP achieved synchronous detection of the morphology, biochemistry, cell cycle, and the DNA content of the cells. CONCLUSION: OTCPP can quickly identify apoptosis and measure the apoptosis rate, thereby unifying qualitative and quantitative analysis.