Selective procedure for the instant identification of cellular apoptosis induced by natural products

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.

Keywords: Apoptosis identification; Fluorescence microscopy; DNA gel electrophoresis; Flow cytometry; Pinus massoniana bark extract; One transient cell processing procedure

Core Tip: According to the global green development hypothesis, the identification of cell apoptosis induced by natural products is a research hotspot. However, routine laboratory identification of apoptosis suffers from experimental errors and poor repeatability. Herein, we explored the simultaneous morphological, biochemical and molecular identification of apoptosis, while reducing experimental errors. We established a protocol to instantly identify apoptosis following natural product treatment, termed one transient cell processing procedure (OTCPP), comprising eleven steps. It could identify apoptosis of cultured LoVo cells in a little as 4 days. The OTCPP procedure is easy, rapid and high efficient, especially for researchers in developing countries.