checked the activity of different signal proteins( p21( WAF), p53, Akt, STAT3) related to deacetylation, cell growth and differentiation by western blot and analysed cell-cycle arrest, if any, by fluorescence-activated cell sorting. After viability assays had been performed with Condurango 30C and with a placebo, the activities of histone de-acetylase( HDAC) enzymes 1 and 2 were measured colorimetrically. While Condurango 30C induced cytotoxicity in HeLa cells in-vitro and reduced HDAC2 activity quite strikingly, it apparently did not alter the HDAC1 enzyme; the placebo had no or negligible cytotoxicity against HeLa cells and could not alter either the HDAC 1 or 2 activity. Data on p21( WAF), p53, Akt, and STAT3 activities and a cell-cycle analysis revealed a reduction in DNA synthesis and G1- phase cell-cycle arrest when Condurango 30C was used at a 2 % dose. In summary, Condurango 30C appeared to trigger key epigenetic events of gene modulation in effectively combating cancer cells, which the placebo was unable to do.
2. Saha S, et al. Calcarea carbonica induces apoptosis in cancer cells in p53-dependent manner via an immuno-modulatory circuit. BMC Complement Altern Med. 2013; 13:230. The authors of this study attempted to evaluate the efficacy of Calcarea carbonica, a homeopathic medicine, as an anti-cancer agent and to delineate the molecular mechanism( s) underlying Calc carb-induced tumour regression via trypan blue dye-exclusion test, flow cytometric, Western blot and reverse transcriptase-PCR techniques. Further, siRNA transfections and inhibitor studies were used to validate the involvement of p53 pathway in Calc carb-induced apoptosis in cancer cells. The results confirmed a significant anti-cancer effect and that Calc carb induced a " two-step " mechanism of the induction of apoptosis in tumour cells, that is,( 1) activation of the immune system of the host; and( 2) induction of cancer cell apoptosis via immunomodulatory circuit in p53-dependent manner by down-regulating Bcl-2: Bax ratio. Bax up-regulation resulted in mitochondrial transmembrane potential loss and cytochrome c release followed by activation of caspase cascade. Knocking out of p53 by RNA-interference inhibited Calc carb-induced apoptosis, thereby confirming the contribution of p53.
3. Saha S, et al. Contribution of the ROS-p53 feedback loop in thujainduced apoptosis of mammary epithelial carcinoma cells. Oncol Rep. 2014; 31( 4): 1589-98. Workers here examined the anti-tumourigenic activity of homeopathically prepared Thuja occidentalis, and the molecular mechanisms underlying thuja-induced apoptosis of functional p53-expressing mammary epithelial carcinoma cells were elucidated. Cells were treated with Thuja or placebo at potencies of 6C, 30C or 200C at different concentrations( 10, 15, 20 and 30 ยต l / ml) for different timepoints( 0, 6, 8, 12, 24, 36 and 48 hours) to select the optimum time required for cell killing. The results showed that Thuja successfully induced apoptosis in functional p53-expressing mammary epithelial carcinoma cells. Abrogation of intracellular reactive oxygen species( ROS), prevention of p53-activation, knockdown of p53 or inhibition of its functional activity significantly abridged ROS generation. Notably, under these conditions, Thuja-induced breast cancer cell apoptosis was reduced, thereby validating the existence of an ROS-p53 feedback loop. Elucidating this feedback loop revealed bi-phasic ROS generation as a key mediator of Thuja-induced apoptosis. The first phase of ROS was instrumental in ensuring activation of p53 via p38MAPK and its nuclear translocation for transactivation of Bax, which induced a second phase of mitochondrial ROS to construct the ROS-p53 feedback loop. Such molecular crosstalk induced mitochondrial changes i) to maintain and amplify the Thuja signal in a positive self-regulatory feedback manner; and ii) to promote the mitochondrial death cascade through cytochrome c release and caspase-driven apoptosis. 28 | vol31 | no1 | JATMS