Studies On The Anti-Tumor And Apoptosis-Inducing Activity Of Betulinic Acid From Semen Ziziphi Spinosae

This paper focused on the study of the in vitro anticancer activity of Betulinic Acid (BetA) from Semen Ziziphi Spinosae, a Traditional Chinese Medicine (TCM).Firstly, crush the Semen Ziziphi Spinosae fruit, degrease with petroleum ether and extract the residue with dichloromethane. Then reflux in dehydrated alcohol, and after dried in vacuo, the precipitate was filtered and recrystallized to yield BetA(1.3‰), which was certificated coincident with those literature spectrum data. MTT assay showed that BetA could inhibit the proliferation, in a dosage dependent manner, of the cells of HT-29,A2780, LS-1747,SMMC-7721,Bcap-37,OVCAR-3,PC-14,K562,MA891 and KB cell lines, in which the IC50 of KB cells was the least.In the study of the BetA's inducing KB cells apoptosis activity, we observed the typical apoptotic morphological changes after treated with BetA. Moreover, TUNEL assay detected the apoptotic DNA cleavage and apoptosis cells were also got by flow cytometer after stained with PI, so that the inducing apoptosis activity of BetA was approved by morphological, biochemical methods and flow cytometry. Besides, it was also found that when incubated with BetA, the Mitochondia membrane voltage of KB cells would decrease dose-dependently, which was the characteristic of the apoptotic cell mitochondria and this further affirmed the apoptotic-inducing effect of BetA on KB cells.With these results, we can come to the conclusion that the BetA from Semen Ziziphi Spinosae had the anticancer activity through inducing tumor cell apoptosis and could be used as a new lead compound for anticancer drugs.