Supporting Information. For

Transcription

1 Supporting Information For A Polyoxometalate-Based Radiosensitization Platform for Treating Hypoxic Tumor by Attenuating Radio-Resistance and Enhancing Radiation Response Yuan Yong 1,, Chunfang Zhang 1, Zhanjun Gu 1, *, Jiangfeng Du 1, Zhao Guo 1, Xinghua Dong, Jiani Xie 1, Guangjin Zhang 3 *, Xiangfeng Liu 1, * *, Yuliang Zhao 1 CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, Institute of High Energy Physics and National Center for Nanoscience Technology of China, Chinese Academy of Sciences, Beijing 19, P.R. China. College of Materials Science and Opto-electronic Technology, University of Chinese Academy of Sciences, Beijing 19, P.R. China. 3 Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 119, P.R. China. College of Chemistry and Environment Protection Engineering, Southwest Minzu University, Chengdu, 11, P.R. China. * Corresponding Authors: zjgu@ihep.ac.cn, liuxf@ucas.ac.cn, zhaoyuliang@ihep.ac.cn, zhanggj@ipe.ac.cn These authors contributed equally. 1 / 1

2 3 5 O Counts Na C W P N Gd Gd W Energy (Kev) Figure S1: The energy-dispersive X-ray (EDS) spectra of the as-prepared GdW nanosphere. / 1

3 (a) (b) (c) Intensity (%) h Diameter (nm) Intensity (%) Diameter (nm) (d) (e) (f) Intensity (%) Diameter (nm) h Intensity (%) 1 Figure S: (a-e) The size distributions of these GdW nanospheres dispersed in DMEM medium with the change of time. (f) The average hydrodynamic diameter of these GdW nanospheres dispersed in DMEM medium as a function of time. h Diameter (%) 7 h Intensity (%) Average Diameter (nm) Diameter (nm) h Time (h) 3 / 1

4 Figure S3: Representative H&E stained images of main organs including heart, liver, spleen, lung and kidney collected from BEL-7 tumor bearing nude mice before and after i.v. injected into male Balb/c mice at a dosage of mg/kg. / 1

5 Hemogram WBC (cells/µl) 1 MCV (fl) PLT (cells/µl) LYM (cells/µl) MID% (%) x1 3 7x1 3 x1 3 5x1 3 x1 3 3x1 3 x1 3 1x Biochemical ALT (U/L) AST (U/L) RBC (cells/µl) MCH (pg) PCT (%) MID (cells/µl) 1 GRN% (%) CRE (µm) HGB (g/dl) MCHC (g/dl) MPV (fl) GRN (cells/µl) URE (mm) HCT (%) RDW (%) PDW (%) LYM% (%) Figure S: Blood chemistry analysis and hematological parameters of mice after i.v. injection with GdW nanosphere ( mg/kg). 5 / 1

6 Liver Function (a) ALT (U/L) PBS GdW Renal Function (c) CREA (µm) X-ray GdW sirna GdW1@CS+X-ray GdW 1 sirna +X-ray PBS GdW X-ray GdW sirna GdW GdW sirna +X-ray (b) AST (U/L) (d) UREA (mm) PBS GdW1@CS PBS GdW X-ray GdW1@CS sirna X-ray GdW1@CS+X-ray GdW1@CS sirna +X-ray GdW sirna GdW GdW sirna +X-ray Figure S5: Hematological parameters of mice after various administrations: (i) PBS only, (ii) GdW nanosphere, (iii) X-ray only, (iv) GdW sirna, (v) GdW (vi) GdW sirna +X-ray. / 1

7 (a) (c) Cell Viability (%) Cell Viability (%) 1 BEL Concentration of GdW sirna (µg/ml) 1 1 Concentration of CoCl (µm) (b) (d) Figure S: In Vitro Cytotoxicity assay of GdW sirna and CoCl under hypoxia. Cytotoxicity of different concentrations of GdW sirna nanosphere to HeLa (a) and BEL-7 cells (b) under hypoxia. Cytotoxicity of HeLa (c) and BEL-7 cells (d) with different concentrations of CoCl. Cell Viability (%) Cell Viability (%) 1 HeLa Concentration of GdW sirna (µg/ml) 1 1 Concentration of CoCl (µm) 7 / 1

8 Figure S7: the sensitizer enhancement ratio (SER) of GdW and GdW sirna in BEL-7 cells under hypoxia. / 1

9 Figure S: In Vitro Colony Formation assay of GdW nanosphere in HeLa cells under hypoxia. (a) Colony Formation assay of HeLa cells incubated with GdW and GdW sirna in a dose-dependent X-ray manner under hypoxia. (b) The corresponding surviving fraction of HeLa cells after different treatments. Inset: the sensitizer enhancement ratio (SER) of GdW and GdW sirna in HeLa cells under hypoxia. (c) Representative γ-hax immufluorescence images of DNA double-strand damage under hypoxia induced by GdW (1 µg ml -1, ml) or/and GdW sirna (1 µg ml -1, ml) or/and X-ray radiation ( Gy), stained with Hoechst and γ-hax for nuclear visualization and DNA fragmentation. (d) The corresponding normalized number of γ-hax under hypoxia after different treatment. Error bars were caculated by SD of three parallel samples. P values were based on the student s test: * P<.5, ** P<.1, *** P<.1. 9 / 1

10 Figure S9: Representative tumor images of different mice: (i) PBS only, (ii) GdW nanosphere, (iii) X-ray only, (iv) GdW sirna, (v) bgdw (vi) GdW1@CS sirna +X-ray. The monitoring cycle is 5 days. GdW nanosphere: mg/ml, µl. X-ray irradiation: 1 Gy. 1 / 1

11 ID%/g PBS h d d 15d 3d heatr liver spleen lung kidney blood Figure S1: Biodistribution of GdW nanosphere at a dosage of mg/kg i.v. injected into male Balb/c mice. 11 / 1

12 Figure S11: H&E staining images. Representative H&E stained images of main organs including heart, liver, spleen, lung and kidney collected from BEL-7 tumor bearing nude mice after various administrations. / 1

13 Figure S: MR and CT imaging in vitro. (a) MR images collected from different concentrations of GdW nanosphere (,.5,.5, 5, 5 mg/ml). (b) CT images collected from different concentrations of iopromide and GdW nanosphere (,.5,.5, 5, 5 mg/ml). 13 / 1

14 Figure S13: MR and CT imaging in BEL-7 tumor-bearing nude mice. (a) MR images collected from BEL-7 tumor-bearing nude mice after intravenous injection (i.v.) of GdW solution (15 mg/ml, µl) at different time point (, 1,, 9, and min). The coresponding signal intensity of MR in liver (b) and tumor (c) before and after i.v. injection of GdW solution. CT images of BEL-7 tumor-bearing nude mice before (d) and after (e) i.v. injection of GdW solution (5 mg/ml, µl) at different orientation: the highlight of details CT images. 1 / 1