摘要
The anti-tumor effects of catechins and gallic acid （GA） in-vitro was investigated in this paper. Fluo-3AM, Calcium-AM （Ca-AM）, 2＇, 7＇ -dichlorofluorescein-diacetate（DCFH-DA）, 4＇ ,6-dia- midino-2- phenylindole （DAPI） and Ca-AM plus colbat were used to characterize intracellular calcium, labile iron pool （LIP）, reac- tive oxygen species （ROS）, nuclei morphology and mitochondrial permeability transition pore （mPTP） opening, respectively. High performance liquid chromatography （HPLC） was used to quanti- tare catechins and GA in the cultural medium. The results indi- cated that each of them showed dose response inhibition of cell growth, provoking nuclei condensation, intracellular calcium ele- vation, mPTP opening, LIP reduction, and cytochrome c （Cyt-C） to release into cytosol. The caspase inhibitors, 2-aminoethox- ydiphenol borate （APB） or Fe3＋ could inhibit lethal effects of GA and （-）-epigallocatechin （EGC）, but failed to affect （-）-epigalloca- techin gallate （EGCG） and （-）-epicatechin gallate （ECG）. Level of ROS presented negative growth while their concentration de- creased in the medium. In conclusion, our findings suggest that viability of RKO decreased because of their good correlation with elevation of calcium and loss of L1P and ROS in cytosol.
The anti-tumor effects of catechins and gallic acid （GA） in-vitro was investigated in this paper. Fluo-3AM, Calcium-AM （Ca-AM）, 2＇, 7＇ -dichlorofluorescein-diacetate（DCFH-DA）, 4＇ ,6-dia- midino-2- phenylindole （DAPI） and Ca-AM plus colbat were used to characterize intracellular calcium, labile iron pool （LIP）, reac- tive oxygen species （ROS）, nuclei morphology and mitochondrial permeability transition pore （mPTP） opening, respectively. High performance liquid chromatography （HPLC） was used to quanti- tare catechins and GA in the cultural medium. The results indi- cated that each of them showed dose response inhibition of cell growth, provoking nuclei condensation, intracellular calcium ele- vation, mPTP opening, LIP reduction, and cytochrome c （Cyt-C） to release into cytosol. The caspase inhibitors, 2-aminoethox- ydiphenol borate （APB） or Fe3＋ could inhibit lethal effects of GA and （-）-epigallocatechin （EGC）, but failed to affect （-）-epigalloca- techin gallate （EGCG） and （-）-epicatechin gallate （ECG）. Level of ROS presented negative growth while their concentration de- creased in the medium. In conclusion, our findings suggest that viability of RKO decreased because of their good correlation with elevation of calcium and loss of L1P and ROS in cytosol.
基金
Supported by Zhejiang Provincial Natural Science Foundation of China(Y3100683)
