Analysis of Relationship between Apoptosis and Changes of Ca~(2+) in HepG-2 Induced by CSA with Laser Scanning Confocal Technology

摘要

Laser scanning confoncal technology was used to study relationship between apoptosis and changes of Ca~(2+) induced by CSA (Capparis spinosa L. total alkaloid, CSA) on human heptocarcinoma cell HepG-2. Killing effects of CSA on human heptocarcinoma cell HepG-2 was measured by MTT method, while morphological observation on HepG-2 cells was completed by fluorescence microscope. Apoptosis of HepG-2 cells induced by CSA was measured by flowcytometry. In addition, change of intracellular Ca~(2+) level of CSA on HepG-2 cells was observed by laser scanning confocal microscope. Results: CSA had obvious cytotoxicity on HepG-2 in a dose-dependent manner, and its IC_(50) was 162.4μg/ml. CSA could induce characteristic apoptosic morphology of HepG-2 cells, and apoptosis percentage was significantly higher than control one. Migration of cells cycle from S phase to G2 phase had been blocked by CSA. Concentration of Ca~(2+) in HepG-2 had been increased by CSA, which was positive correlation with drug dosage. CSA had obvious effects of killing and inducing apoptosis on human heptocarcinoma cell HepG-2, and overload of Ca~(2+) might be involved in the se events.