Paclitaxel

2-mo-old C57BL6/J mice (18–25 g) of both sexes were injected i.p. with 20 mg/kg paclitaxel (Bristol-Myers Squibb) every other day (days 1, 3, 5, and 7, for a total of four injections), while 2 mg/kg 8-Br–cADPR was injected after every two paclitaxel injections (days 4 and 8, for a total of two injections). paclitaxel was prepared in one part vehicle (1:1 vol/vol Cremophor EL [EMD Millipore] and 200-proof ethanol) and two parts sterile saline (UPS) and injected at 10 µl/g. Control mice were injected with one part vehicle and two parts saline. 8-Br–cADPR (Sigma-Aldrich) was prepared in sterile saline and injected at 10 µl/g, and control mice were injected with saline. Mice were habituated for 2 d, and baseline behavioral performance was assessed over the next 2 d and averaged. The first paclitaxel injection (day 1) was given 3 d later, and mice were behaviorally tested 9 d after the final injection (day 17). Noxious mechanosensation threshold was assayed as described previously (Pease-Raissi et al., 2017 (link)) using Von Frey filaments (0.008–1.4 g). Withdrawal threshold was determined to be the applied force at which the animal withdrew the stimulated paw on at least 2 of 10 applications.

This was a retrospective analysis of patients with HER2-negative MBC who received treatment with bevacizumab (Fritz Hoffmann-La Roche, Basel, Switzerland; 10 mg/kg on days 1 and 15, every 28 days) plus paclitaxel (Bristol-Myers Squibb, New York, NY, USA; 80 mg/m² on days 1, 8, and 15, every 28 days) as first-line therapy, second-line therapy, or subsequent lines, including those who received bevacizumab continuation therapy after completion of paclitaxel, at La Paz University Hospital between August 2007 and October 2012. All analyses were approved by the local ethics committee.

Avelumab (designated as α-PD-L1) and M7824 were generously provided by EMD Serono (Billerica, MA). The following inhibitors were used: actinomycin D (RNA synthesis inhibitor, Sigma-Aldrich, St. Louis, MO), SD-208 (TGF-βRI inhibitor, R&D Systems), SIS3 (Smad3 inhibitor, Tocris Bioscience, Bristol, UK), wortmannin (PI3K inhibitor, EMD Millipore, Darmstadt, Germany), U0126 (MEK1/2 inhibitor, Cell Signaling Technology, Danvers, MA), SB203580 (p38 MAPK inhibitor, Selleckchem, Houston, TX), BMS-345541 (NF-κB inhibitor, Sigma-Aldrich), and S31–201 (STAT3 inhibitor, Sigma-Aldrich). The following chemotherapy drugs were used: docetaxel (Sanofi-Aventis, Paris, France), paclitaxel (Bristol-Myers Squibb, New York, NY), and gemcitabine (Tocris Bioscience).

Paclitaxel was purchased from Bristol-Myers Squibb (New York, NY, USA). A Cell Counting Kit-8 (CCK-8) was obtained from Dojindo Molecular Technologies (Kumamoto, Japan). The Protease Inhibitor Cocktail was provided by Roche (Basel, Switzerland). Mouse anti-human COX4 antibody was obtained from Molecular Probes (Eugene, OR, USA). Goat anti-human lamin B and mimitin antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse anti-human flotillin-1 and rabbit anti-human β-actin antibodies were obtained from eBioscience (San Diego, CA, USA). Rabbit anti-human 14-3-3 ζ/δ antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively. The electrochemiluminescence kit was purchased from Thermo Scientific (Boston, MA, USA).