Additional file 1 of Two nanoformulations induce reactive oxygen species and immunogenetic cell death for synergistic chemo-immunotherapy eradicating colorectal cancer and hepatocellular carcinoma
Creators: name: Guo, Jianfeng
name: Yu, Zhuo
name: Sun, Dandan
name: Zou, Yifang
name: Liu, Yun
name: Huang, Leaf
Publisher: name: figshare
Year: 2021
Subjects: Space Science
Medicine
Chemical Sciences not elsewhere classified
Biological Sciences not elsewhere classified
Mathematical Sciences not elsewhere classified
Cancer

description: Additional file 1 Fig. S1. The physicochemical characterization of non-targeted Nano-FdUMP. A) TEM image (bar = 100 nm). B) Size distribution (~ 38 nm, polydispersity index ≈ 0.3) and surface charge (~ 5 mV). C) The in vitro release of fluorine drug from nanoprecipitates in pH = 5.5 and 7.4 (n = 4). d) No significant aggregation was caused in 10% serum-containing medium up to 12 h at 37 °C. Fig. S2. Toxicity of Nano-FdUMP in healthy BALB/C mice. A) The body weight over a 35-day period following treatment of PBS and Nano-FdUMP containing 5, 10, 25 and 50 mg/kg FdUMP on Day 1, 3 and 5. B) The overall condition of animals (n = 5) based on body condition scoring [BCS, IACUC Guidelines along with other criteria (e.g., hunched posture, ruffled hair coat, and reluctance to move)]. At the endpoint, the number of animals compliant with BCS index was presented. Results of non-targeted Nano-FdUMP were similar to those observed in targeted counterpart (Data not shown). Fig. S3. Blood circulation of non-targeted Nano-FdUMP in orthotopic CRC and HCC mouse models. Following i.v. injection, the concentration of fluorine drug on different time points was plotted (n = 4). Results showed that non-targeted Nano-FdUMP demonstrated similar blood circulation recorded by targeted counterpart (Fig. 3a). Fig. S4. Therapeutic efficacy of Nano-FdUMP in orthotopic CRC and HCC mouse models. Following treatment schedule as described in Figs. 5 and 6, Nano-FdUMP at doses of 10 and 25 mg/kg FdUMP achieved significantly improved antitumor efficacy as compared to PBS and 5-Fu at 50 mg/kg (n = 5, * p &lt; 0.05 and p &lt; 0.01). Fig. S5. Therapeutic efficacy of Nano-FdUMP with/without AEAA at dose of 10 mg/kg FdUMP in orthotopic CRC and HCC mouse models. Following treatment schedule as described in Figs. 5 and 6, non-targeted Nano-FdUMP could not slow down tumor growth as compared to PBS, but AEAA-targeted Nano-FdUMP achieved significantly improved antitumor efficacy than PBS and non-targeted Nano-FdUMP (n = 5, * p &lt; 0.05). Fig. S6. Immunofluorescent staining of tumors on Day 24 (as described in Fig. 5) (cleaved caspase 3 = green; nuclei = blue) to determine apoptosis (n = 3, ** p &lt; 0.01, relative to Nano-Folox/5-Fu). Fig. S7. Rechallenge studies. A) Orthotopic CRC mice (BALB/C) were treated with Nano-FdUMP/Nano-Folox as described in Fig. 5, and one month after tumor disappearance, two flanks of mice were respectively rechallenged with 4 T1 and CT26-FL3 cells (1 × 106 cells per mouse) (n = 4). B) Orthotopic HCC mice (C57BL/6) were treated with Nano-FdUMP/Nano-Folox as described in Fig. 6, and one month after tumor disappearance, two flanks of mice were respectively rechallenged with B16 and Hepa1–6-Luc cells (1 × 106 cells per mouse) (n = 4). Tumor volume was calculated using the formula a2b(π/6), where a is the minor diameter of the tumor and b is the major diameter perpendicular to diameter a. Results confirmed that Nano-FdUMP/Nano-Folox could induce tumor-specific memory response. Fig. S8. Immunofluorescent staining of tumors on Day 23 (as described in Fig. 6) (cleaved caspase 3 = green; nuclei = blue) to determine apoptosis (n = 3, ** p &lt; 0.01, relative to Nano-Folox/5-Fu). Fig. S9. Toxicity studies of two nanoformulations in A) BALB/C and B) C57BL/6 mice. The body weight over a 35-day period following treatment of PBS and combination of two nanoformulations (Nano-Folox containing 1.5 mg/kg platinum drug was i.v. injected into mice on Day 1, 3 and 5. Eight hours post injection, Nano-FdUMP containing 10 mg/kg fluorine drug was i.v. injected into mice). Results show that no significant change was found in body weight and hematological/liver/kidney functions following treatment of two nanoformulations as compared to PBS (n = 5). Fig. S10. Immunofluorescent staining of tumors on Day 12 (as described in Fig. 7) (cleaved caspase 3 = green; nuclei = blue) to determine apoptosis (n = 3, ** p &lt; 0.01, relative to Nano-FdUMP/Nano-Folox). Table S1. Antibodies used in the study. Table S2. Primers used for RT-PCR in the study.