Anti-cancer Effect of Andrographis paniculata, Ziziphus spina-christi,and Vernonia extensa and its Combination with Piperine Free Piper nigrum Extract on Cancer Cell Lines

Abstract

Andrographis paniculata, Ziziphus spina-christi, Vernonia extensa, and Piper nigrum have biological activities in anti-cancer activity. This study was carried out to investigate the anti-cancer activity of three plants individually and the interaction of combination among three plants leaves and PFPE as a novel anti-cancer regimen as well as to analyze the apoptosis and multi-caspase activities induced by the combination. The study results demonstrated that dichloromethane extract of A. paniculata leaves exhibited anti-proliferation on colorectal cancer cell lines (HT-29 and SW-620) with an ICso values of 8.93 # 0.52 g/ml and 7.49 $ 0.04 4g/ml, respectively. The GC-MS analysis suggests that major volatile constituents group of this extract of A. paniculata leaves are alcohols group (1-heptatriacotanol), and steroids (androsta-1,4-dien-3-one, 6,17-dihydroxy-, (63,17ß) and y-sitosterol). The dichloromethane extract of Z. spina-christi and V. extensa showed the highest cytotoxicity on MCF-7 and A2780 cells with an ICso values of 13.35 $ 0.30 ug/ml and 14.64 $ 1.51 ug/ml, respectively. The major volatile compounds group of Z. spina-christi are including fatty acids (palmitic acid, linolenic acid, and linoleic acid), terpenes (neophytadiene), and fatty alcohols (n-tetracosanol-1). The dichloromethane extract of V. extensa exhibited the highest cytotoxicity in MCF-7 and A2780 cells with ICso at 15.58 ÷ 1.81 ug/ml and 10.08 0.04 pg/ml, respectively. The major volatile compounds of dichloromethane extract of V. extensa are fatty acids (palmitic acid, linolenic acid, and linoleie acid), and terpenes (neophytadiene and phytol). Thereafter, the combination study was carried out using dichloromethane extract of A. paniculata, Z. spina-christi, and V. extensa leaves, combined with PFPE each dichloromethane extract at various ratio (ICso: IC so; ICso:0.51Cso; 0.5ICso:ICso, and 0.5ICso:ICso) (where ICso as a baseline). The combination among the dichloromethane extract of V. extensa leaves and PFPE showed a synergistic interaction on MCF-7 cell lines with a CI value of 0.34 ÷ 0.06; 0.36 $ 0.14, and 0.44 # 0.14, respectively. As a further investigation, the combination of dichloromethane extract of V. extensa and PFPE was tested in apoptosis and multi- caspase activities assessed by Annexin V-FITC and Multi-caspase assays. The results showed that combination of dichloromethane extract of V. extensa leaves with PFPE at ratio ICso:ICso induced caspases cascade and leading to apoptosis with time-dependent on MCF-7 cell lines. In conclusions, dichloromethane extract of A. paniculata, Z. spina-christi, and V. extensa represented a potential anti-cancer treatment in the individual extract. The combination of dichloromethane extract of V. extensa and PFPE could be a promising anti-cancer regimen in combating breast cancer cell lines.