Fig. 7: Inhibition of OXPHOS and mitochondrial translation significantly impairs spheroid formation in vitro and tumor progression in vivo.

a–i Representative images and quantification of A4 spheroids exposed to various drugs and their combinations (Panels with red star denotes the antibiotic drugs Doxycycline and Erythromycin that most significantly inhibited spheroid formation over other the drugs, and hence were subsequently used for in vivo evaluation); a-ii Dot plot representing the number of spheroids under different drug treatment measured on Day14 (post cell seeding); b Schematic representation of drug regimen used in the study; c-i Representative A4 tumors harvested on Day 21 of different drug regimens; c-ii Line graph indicating A4 tumor volumes measured on Day 0,7,14 and 21 during treatment of various drugs (single and in combination; Day 0 corresponds to the initiation of treatment, which occurs 14 days post subcutaneous tumor cell inoculation); c-iii. Bar-graph indicating the anti-tumor efficacy of various drug combinations, Y-axis represents the tumor inhibitory score (where control represent a score of “1”); d-i Line graph depicting A4 tumor volumes over the course of treatment with the ETC complex I inhibitor metformin, administered either as a monotherapy or in combination with other therapeutic agents. Day 0 marks the initiation of treatment. Asterisks of different colors denote p-value significance (student’s t test) between each treatment group and the vehicle control on the corresponding day. The dotted line denotes p-vlaue significance among the various drug regimens on Days 7, 14, and 21, as assessed by one-way ANOVA. The solid line indicates the statistical significance of the comparison between Metformin monotherapy and the combination therapy comprising Metformin, Erythromycin, and Paclitaxel (Metformin + Ery + Pax); d-ii Representative A4 tumors harvested on 21st day after different drug regimens. *p < 0.05, **p < 0.01, and ***p < 0.001.