Figure 7

Calcarea carbonica induces T cell-mediated apoptosis of primary mammary tumor. (A) T cells isolated from patient’s peripheral circulation were primed with media-/placebo-/calcarea carbonica-treated tumor supernatant for 72 h and then co-cultured with primary mammary tumor for 48 h. In parallel, primary mammary tumor cells was directly exposed to placebo-/calcarea carbonica for 48 h in the absence of T cells. Tumor cell apoptosis was then scored by Annexin-V-PE/7-AAD-positivity and represented graphically. (B) T cells isolated from normal and cancer patient’s peripheral blood were co-cultured with control mammary tissue and tumor mammary tissue explants, respectively for 48 h and percent T cell apoptosis was scored by Annexin-V-PE/7-AAD-positivity. (C) The same experimental set was analysed for percentage of CD4⁺ and CD8⁺ T cells flow cytometrically. (D) Lysates of primary breast cancer cells co-cultured with or without placebo-/calcarea carbonica-primed T cells of same patient’s blood were Western blotted for the analysis of p53, Bax, Bcl-2 and active-caspase-3. α-Actin was used as loading control. *p < 0.05 and **p < 0.001 when compared with respective control/mammary tissue explant sets and placebo/drug-treated sets.