Anti-tumor Effects Of Chinese Ephedra Herb And Henbane Seed On Human Breast Cancer SKBR3Cells

Objective:The present study was designed to investigate the effects of ephedrine and anisodamine on proliferation, distribution of cell cycle and apoptosis of human breast cancer SKBR3cells and explore the underlying antitumor mechanism of Chinese ephedra herb and henbane seed, so as to provide a theoretical basis for antitumor effect of Chinese herbal medicine.Methods:Human breast cancer SKBR3cells,5X103cells/well or1X105cells/well were seeded in24-well or6-well plates.24hours later, the medium was changed and the cells were exposed to the indicated reagents or vehicle for the indicated time. Groups were divided as follow:(1) Ephedrine experimental groups:control group, low-dose ephedrine group (1μg/ml), medium-dose ephedrine group (10μg/ml), high-dose ephedrine group(100μg/ml), low-dose epinephrine group (0.05μg/ml), medium-dose epinephrine group (0.5μg/ml) and high-dose epinephrine group (5μg/ml);(2) Anisodamine experimental groups:control group, low-dose anisodamine group (0.2μg/ml), medium-dose anisodamine group (2μg/ml), high-doses anisodamine group (20μg/ml), low-dose atropine group (0.015μg/ml), medium-dose atropine group (0.15μg/ml) and high-dose atropine group (1.5μg/ml). Cell morphological changes were observed under light microscope. The antiproliferative activity and cell-survival rates were determined by trypan blue counting method. Cell-apoptosis and cell-cycle distribution were investigated using flow cytometry.Results:1Ephedrine and anisodamine could inhibite proliferation of SKBR3, and the highest inhibition rates were42.88%and28.57%, respectively.2There were no obvious cytotoxicity to SKBR3cells in both ephedrine and anisodamine groups.3Compared with control group, the number of SKBR3cells in Go/G1Phase in both ephedrine and anisodamine group increased gradually, while the number of SKBR3cells in S phase and G2/M Phase decreased and proliferation index also tapered. The distribution of cell cycle in both high-dose ephedrine group and high-dose anisodamine group had significant difference compared with corresponding control group (ephedrine:Go/Gi phase84.010±0.015VS70.705±0.795P<0.01,S phase6.655±0.060VS9.120±0.254P<0.01,G2/M phase9.330±0.069VS20.175±0.713P<0.01; anisodamine:Go/G1phase63.305±0.632VS53.530±0.242P<0.01,S phase28.760±0.727VS34.550±0.283P<0.01,G2/M phase7.935±0.095VS11.92±0.525P<0.01).4Ephedrine could weakly induce apoptosis of SKBR3cells, while anisodamine didn’t show the effect of inducing apoptosis of SKBR3cellsConclusions:1Ephedrine and anisodamine could inhibite proliferation of SKBR3cells.2Both ephedrine and anisodamine have no obvious cytotoxicity to SKBR3cells.3Both ephedrine and anisodamine can arrest SKBR3cells cycle in GO/G1stage.4Ephedrine can weakly induce apoptosis of SKBR3cells, while anisodamine might not induce apoptosis of SKBR3cells.5Ephedrine and anisodamine can function in β-adrenergic receptor and cholinergic receptor, thereby mediating the sympathetic and parasympathetic nervous system in a new balance, and then change the level of intracellular cAMP and affect the function of tumor cells.6The possible anti-tumor mechanism of Chinese ephedra herb and henbane seed maybe that ephedrine and anisodamine can function in β-adrenergic receptor and cholinergic receptor, thereby mediating the sympathetic and parasympathetic nervous system in a new balance.This balance led to changes in the level of intracellular cAMP, thereby affect the distribution of cell cycle and inhibite proliferation of SKBR3cells. But the specific mechanism need further study.