Abstract C108: Therapeutic potential of anticancer polyherbal formulations.

Field: Pharmacology, Toxicology and Pharmaceutics (Life Sciences)

Abstract Background: Cancer is a leading cause of death worldwide and poses a huge health challenge (1). The global burden of cancer continues to increase despite advances in cancer therapeutics. There is renewed interest in plant-based drugs and growing appreciation of the use of polyherbal formulations (2). Multitargeted polyherbal formulations provide synergistic or additive preventive effects when the agents are combined could theoretically permit the administration of the lowest active dose of each agent and therefore lower the potential for adverse side effects. Hepatocellular carcinoma (HCC) is one of the most prevalent malignancies and generally considered resistant to chemo- and radiotherapies. Resisting cell death and angiogenesis are two important hallmarks of cancer cells. Therefore, apoptosis and blocking of angiogenesis are crucial therapeutic approaches. The present study aims to evaluate the anticancer potential of four polyherbal formulations on human hepatocellular carcinoma (Hep-G2) cells and their antiangiogenic potential on chick-chorioallantoic membrane. Materials and Methods: The following were the studies carried out: (i) cytotoxicity using MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay; (ii) apoptosis detection using Annexin V-FITC Propidium Iodide staining and flow cytometry [the frequency of annexin-positive cells (apoptotic) was determined using BD FACSDiva software]; (iii) DNA fragmentation and chromatin condensation evaluated with Hoechst 33258 staining and flourescent microscopy; and (iv) evaluation of atiangiogenic potential the formulations using ex-ovo chick Chorio-Allantoic Membrane (CAM) assay. Results: All four formulations (VK, MK, KG, and GTK) showed significant cytotoxicity with inhibitory concentration (IC50) of 48.6, 57.0, 41.7, and 48.3 µg/ml, respectively at 24h and 32.4, 31.2, 37.5, 29.9 µg/ml, respectively at 48h of treatment. Maximum apoptosis was observed at concentration of 100 µg/ml in HepG2 cells after 48 h of treatment. Treatment induced apoptosis (cumulative) was 98.8% (VK), 94% (MK), 91.7% (KG), and 97.9% (GTK) at 100 µg/ml, in comparison to 88.6% for paclitaxel (10 µg/ml). Apoptosis was further confirmed by the observation of DNA fragmentation and chromatin condensation in fluorescent microscopy. The formulation VK showed maximum anti-neoplastic activity followed by GTK, MK, and KG. It is interesting to note that the cell death pattern observed with the polyherbal formulations is comparable to that of the synthetic single molecule drug paclitaxel. The microscopic evaluation and morphometric image analysis of area of vascularization revealed that all four formulations showed significant inhibition of neo-vascularization. Maximum anti-angiogenic activity was observed at 100 µg/ml. Inhibition of vascularization observed at 100 µg/ml was 36.6 ± 3.5 mm2 (VK), 30.9 ± 7.0 mm2 (MK), 32.6 ± 6.6 mm2 (KG), and 33.2 ± 4.3 mm2 (GTK). On the other hand, thalidomide resulted in 36.4 ± 3.6 mm2 at 10 µg/ml. Conclusions: All four classic polyherbal formulations showed significant anticancer and antiangiogenic activity compared to chemothereputic drugs. However, VK and MK showed the maximum anticancer and antiangiogenic activity, respectively. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C108. Citation Format: Gaurav Sharma, Rama Jayasundar, Shyam S. Chauhan, Thirumurthy Velpandian. Therapeutic potential of anticancer polyherbal formulations. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C108.