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 µ 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 .
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28 | vol31 | no1 | JATMS