Ext web page.)Ebert et al. Molecular cancer 2014, 13:265 http:molecular-cancercontent131Page 7 of
Ext page.)Ebert et al. Molecular Cancer 2014, 13:265 http:molecular-cancercontent131Page 7 of(See figure on preceding page.) Figure 3 Cell viability and caspase 37 activity in breast cancer cells co-treated with probenecid and bisphosphonates. Cell viability (A-L) and caspase 37 activity (M-X) was determined in MCF-7, T47D and MDA-MB-231 breast cancer cells right after treatment with ZA (zoledronic acid), RIS (risedronate), IBN (ibandronate), ALN (alendronate) alone and in mixture with probenecid. All data are expressed as signifies of six diverse measure points of three independent experiments as % of controls SEM. Significances have been calculated using the Mann Whitney U test (p 0.005, p 0.05). BP: bisphosphonate, black line; Prob: probenecid, dotted line probenecid co-treatment.by intracellular BP effects, e.g. IPPApppI accumulation and inhibition of protein prenylation. We analyzed if other BP are also capable to modulate KLF2 expression in breast cancer cells and if probenecid can improve the observed effects. In MCF-7 cells ZA induced a 13-fold enhance in KLF2 expression, which was further enhanced by Prob cotreatment (32.4-fold expression) when compared with untreated controls (Table 1). Additive effects of Prob had been also observed when employing ALN. The bisphosphonate alone induced KLF2 expression by the factor five.eight using a additional stimulatory effect of Prob co-treatment to a 36.1-fold induction. IBN alone had no influence on KLF2 expression butA7induc on by Prob5 4 3 two 1 0 ZA RIS MCF-7 IBN ALNIPP ApppIwith Prob co-stimulation the expression of KLF2 enhanced 6.1-fold in contrast to RIS, where no co-stimulatory impact of Prob on the absent RIS effect might be observed. In MDA-MB-231 cells ZA and IBN had no substantial influence on KLF2 expression but Prob was able to enhance KLF2 expression 5.1-fold in ZA and four.BRD7 drug 8-fold in IBN costimulatory experiments. RIS alone improved KLF2 expression by the aspect 3.five but Prob co-treatment abandoned the effect to a non-significant expression. No effect was noticed when ALN was utilised, independent of Prob cotreatment. In T47D cells no additive effect of Prob was detectable. ZA elevated KLF2 expression three.0 fold but Prob had no additive impact (two.6-fold expression) just as with regards to IBN, exactly where both IBN and IBNProb treated samples showed an upregulation of KLF2 by the issue 2.two. RIS alone increased KLF2 expression by the element 2.1 but no substantial enhancement was detectable in RISProb treated cells. ALN alone or the combination Caspase 6 custom synthesis ALNProb didn’t influence the expression of KLF2.Breast cancer cells express probenecid sensitive channels and transporters BP onlyThe expression in the pyrophosphate channel ankylosis protein homolog (ANKH), the hemichannel protein pannexin 1 (PANX1), multidrug resistance linked protein 1 (ABCC1) and solute carrier family members 22 (organic anionTable 1 Effects of co-treatment of breast cancer cell lines with probenecid and bisphosphonates around the expression of KLFKLF2 expression MCF-7 13.0 (two.3-60.eight) 32.four (5.8-198.five) 1.six (0.3-10.1) four.two (0.7-35.9) 2.4 (0.5-15.two) six.1 (0.8-31.7) five.eight (1.2-33.4) 36.1 (9.7-141.4) 1.0 (0.3-5.0) T47D three.0 (1.2-7.six) 2.6 (1.0-6.7) 2.1 (1.0-3.7) 1.7 (0.7-4.7) two.2 (0.9-4.9) 2.two (0.9-5.9) two.0 (0.8-5.5) 1.8 (0.8-5.six) 1.0 (0.8-1.3) MDA-MB-231 three.1 (0.6-16.0) 5.1 (0.7-25.six) 3.five (0.6-18.eight) three.4 (0.5-19.two) two.4 (0.3-17.three) four.eight (0.7-28.four) 1.4 (0.2-11.4) three.two (0.4-31.1) 1.3 (0.1-9.four)B7induc on by Prob5 4 three 2 1 0 ZA RIS T47D IBN IPP ApppIZA 20 M ZA Prob RIS 50 M RIS Prob IBN 50 M I.
