Au@SiO_2 @CuInS_2–ZnS/Anti-AFP fluorescent probe improves HCC cell labeling
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摘要
Background: Clear tumor imaging is essential to the resection of hepatocellular carcinoma(HCC). This study aimed to create a novel biological probe to improve the HCC imaging. Methods: Au nano-flower particles and CuInS_2 –Zn S core-shell quantum dots were synthesized by hydrothermal method. Au was coated with porous SiO_2 and combined with anti-AFP antibody. HCC cell line HepG2 was used to evaluate the targeting efficacy of the probe, while flow cytometry and MTT assay were used to detect the cytotoxicity and bio-compatibility of the probe. Probes were subcutaneously injected to nude mice to explore light intensity and tissue penetration. Results: The fluorescence stability of the probe was maintained 100% for 24 h, and the brightness value was 4 times stronger than that of the corresponding CuInS_2 –Zn S quantum dot. In the targeting experiment, the labeled HepG2 emitted yellow fluorescence. In the cytotoxicity experiments, MTT and flow cytometry results showed that the bio-compatibility of the probe was fine, the inhibition rate of HepG2 cell with 60% Cu-QDs/Anti-AFP probe and Au-QDs/Anti-AFP probe solution for 48 h were significantly different(86.3% ±7.0% vs. 4.9% ±1.3%, t = 19.745, P < 0.05), and the apoptosis rates were 83.3% ±5.1% vs. 4.4% ±0.8%( P < 0.001). In the animal experiment, the luminescence of the novel probe can penetrate the abdominal tissues of a mouse, stronger than that of CuInS_2 –ZnS quantum dot. Conclusions: The Au@SiO_2 @CuInS_2 –ZnS/Anti-AFP probe can targetedly recognize and label HepG2 cells with good bio-compatibility and no toxicity, and the strong tissue penetrability of luminescence may be helpful to surgeons.
Background: Clear tumor imaging is essential to the resection of hepatocellular carcinoma(HCC). This study aimed to create a novel biological probe to improve the HCC imaging. Methods: Au nano-flower particles and CuInS_2 –Zn S core-shell quantum dots were synthesized by hydrothermal method. Au was coated with porous SiO_2 and combined with anti-AFP antibody. HCC cell line HepG2 was used to evaluate the targeting efficacy of the probe, while flow cytometry and MTT assay were used to detect the cytotoxicity and bio-compatibility of the probe. Probes were subcutaneously injected to nude mice to explore light intensity and tissue penetration. Results: The fluorescence stability of the probe was maintained 100% for 24 h, and the brightness value was 4 times stronger than that of the corresponding CuInS_2 –Zn S quantum dot. In the targeting experiment, the labeled HepG2 emitted yellow fluorescence. In the cytotoxicity experiments, MTT and flow cytometry results showed that the bio-compatibility of the probe was fine, the inhibition rate of HepG2 cell with 60% Cu-QDs/Anti-AFP probe and Au-QDs/Anti-AFP probe solution for 48 h were significantly different(86.3% ±7.0% vs. 4.9% ±1.3%, t = 19.745, P < 0.05), and the apoptosis rates were 83.3% ±5.1% vs. 4.4% ±0.8%( P < 0.001). In the animal experiment, the luminescence of the novel probe can penetrate the abdominal tissues of a mouse, stronger than that of CuInS_2 –ZnS quantum dot. Conclusions: The Au@SiO_2 @CuInS_2 –Zn S/Anti-AFP probe can targetedly recognize and label HepG2 cells with good bio-compatibility and no toxicity, and the strong tissue penetrability of luminescence may be helpful to surgeons.
Background: Clear tumor imaging is essential to the resection of hepatocellular carcinoma(HCC). This study aimed to create a novel biological probe to improve the HCC imaging. Methods: Au nano-flower particles and CuInS_2 –Zn S core-shell quantum dots were synthesized by hydrothermal method. Au was coated with porous SiO_2 and combined with anti-AFP antibody. HCC cell line HepG2 was used to evaluate the targeting efficacy of the probe, while flow cytometry and MTT assay were used to detect the cytotoxicity and bio-compatibility of the probe. Probes were subcutaneously injected to nude mice to explore light intensity and tissue penetration. Results: The fluorescence stability of the probe was maintained 100% for 24 h, and the brightness value was 4 times stronger than that of the corresponding CuInS_2 –Zn S quantum dot. In the targeting experiment, the labeled HepG2 emitted yellow fluorescence. In the cytotoxicity experiments, MTT and flow cytometry results showed that the bio-compatibility of the probe was fine, the inhibition rate of HepG2 cell with 60% Cu-QDs/Anti-AFP probe and Au-QDs/Anti-AFP probe solution for 48 h were significantly different(86.3% ±7.0% vs. 4.9% ±1.3%, t = 19.745, P < 0.05), and the apoptosis rates were 83.3% ±5.1% vs. 4.4% ±0.8%( P < 0.001). In the animal experiment, the luminescence of the novel probe can penetrate the abdominal tissues of a mouse, stronger than that of CuInS_2 –ZnS quantum dot. Conclusions: The Au@SiO_2 @CuInS_2 –Zn S/Anti-AFP probe can targetedly recognize and label HepG2 cells with good bio-compatibility and no toxicity, and the strong tissue penetrability of luminescence may be helpful to surgeons.
引文
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[15] Kamkaew A, Sun H, England CG, Cheng L, Liu Z, Cai W. Quantum dot-NanoLuc bioluminescence resonance energy transfer enables tumor imaging and lymph node mapping in vivo. Chem Commun(Camb)2016;52:6997-7000.
[16] Jiang TT, Yin NQ, Liu L, Song J, Huang QP, Zhu LX, et al. Au nanoflower@Si02@CdTe/CdS/ZnS quantumdot multi-functional nanoprobe for photothermal treatment and cellular imaging. RSC Adv 2014;4:23630-23636.
[17] Sugawa K, Tamura T, Tahara H, Yamaguchi D, Akiyama T, Otsuki J, et al. Metal-enhanced fluorescence platforms based on plasmonic ordered copper arrays:wavelength dependence of quenching and enhancement effects. ACS Nano 2013;7:9997-10010.
[18] Yuan L, Zhu J, Ren Y, Bai S. Sectional area-dependent plasmonic shifting in the truncated process of silver nanoparticles:from cube to octahedron. J Nanopart Res 2011;13:6305-6312.
[19] Sabaeian M, Khaledi-Nasab A. Size-dependent intersubband optical properties of dome-shaped InAs/GaAs quantum dots with wetting layer. Appl Opt2012;51:4176-4185.
[20] Naumova AV, Modo M, Moore A, Murry CE, Frank JA. Clinical imaging in regenerative medicine. Nat Biotechnol 2014;32:804-818.
[21] Hong GS, Antaris AL, Dai HJ. Near-infrared fluorophores for biomedical imaging. Nature Bio Eng 2017;1:0010.
[22] Hyun H, Owens EA, Wada H, Levitz A, Park G, Park MH, et al. Cartilage-specific near-infrared fluorophores for biomedical imaging. Angew Chem Int Ed Engl2015;54:8648-8652.
[23] Legant WR, Shao L, Grimm JB, Brown TA, Milkie DE, Avants BB, et al. High--density three-dimensional localization microscopy across large volumes. Nat Methods 2016;13:359-365.
[24] Cui W, Li J, Zhang Y, Rong H, Lu W, Jiang L. Effects of aggregation and the surface properties of gold nanoparticles on cytotoxicity and cell growth.Nanomedicine 2012;8:46-53.
[25] Kolny-Olesiak J, Weller H. Synthesis and application of colloidal CuInS2 semiconductor nanocrystals. ACS Appl Mater Interfaces 2013;5:12221-12237.
[26] Chen Y, Li S, Huang L, Pan D. Low-cost and gram-scale synthesis of watersoluble Cu-In-S/ZnS core/shell quantum dots in an electric pressure cooker.Nanoscale 2014;6:1295-1298.
[27] Shen S,Zhao L, Zhou Z, Guo L. Enhanced photocatalytic hydrogen evolution over Cu-doped ZnIn2S4 under visible light irradiation. J Phys Chem C
[2] Zuo TT, Zheng RS, Zhang SW, Zeng HM, Chen WQ. Incidence and mortality of liver cancer in China in 2011. Chin J Cancer 2015;34:508-513.
[3] Zhao YJ, Ju Q, Li GC. Tumor markers for hepatocellular carcinoma. Mol Clin Oncol 2013;1:593-598.
[4] Ariff B, Lloyd CR, Khan S, Shariff M, Thillainayagam AV, Bansi DS, et al. Imaging of liver cancer. World J Gastroenterol 2009;15:1289-1300.
[5] Kokudo N, Ishizawa T. Clinical application of fluorescence imaging of liver cancer using indocyanine green. Liver Cancer 2012;1:15-21.
[6] Liu D, Wang L, Ma S, Jiang Z, Yang B, Han X, et al. A novel electrochemiluminescent immunosensor based on CdS-coated ZnO nanorod arrays for HepG2cell detection. Nanoscale 2015;7:3627-3633.
[7] Shi Y, Pramanik A, Tchounwou C, Pedraza F, Crouch RA, Chavva SR, et al. Multifunctional biocompatible graphene oxide quantum dots decorated magnetic nanoplatform for efficient capture and two-photon imaging of rare tumor cells.ACS Appl Mater Interfaces 2015;7:10935-10943.
[8] Shao D, Li J, Pan Y, Zhang X, Zheng X, Wang Z, et al. Noninvasive theranostic imaging of HSV-TK/GCV suicide gene therapy in liver cancer by folate-targeted quantum dot-based liposomes. Biomater Sci 2015;3:833-841.
[9] Yang MY, Hong J, Zhang Y, Gao Z, Jiang TT, Song J, et al. Bio-compatibility and cytotoxicity studies of water-soluble CuInS2-ZnS-AFP fluorescence probe in liver cancer cells. Hepatobiliary Pancreat Dis Int 2016;15:406-411.
[10] Song J, Ma C, Zhang W, Li X, Zhang W, Wu R, et al. Bandgap and structure engineering via cation exchange:from binary Ag2S to ternary AgInS2, quaternary AgZnInS alloy and AgZnInS/ZnS core/shell fluorescent nanocrystals for bioimaging. ACS Appl Mater Interfaces 2016;8:24826-24836.
[11] Ballou B, Lagerholm BC, Ernst LA, Bruchez MP, Waggoner AS. Noninvasive imaging of quantum dots in mice. Bioconjug Chem 2004;15:79-86.
[12] Choi HS, Liu W, Misra P, Tanaka E, Zimmer JP, Itty Ipe B, et al. Renal clearance of quantum dots. Nat Biotechnol 2007;25:1165-1170.
[13] Zheng X, Wang X, Mao H, Wu W, Liu B, Jiang X. Hypoxia-specific ultrasensitive detection of tumours and cancer cells in vivo. Nat Commun 2015;6:5834.
[14] Wada H, Hyun H, Vargas C, Gravier J, Park G, Gioux S, et al. Pancreas-targeted NIR fluorophores for dual-channel image-guided abdominal surgery. Theranostics 2015;5:1-11.
[15] Kamkaew A, Sun H, England CG, Cheng L, Liu Z, Cai W. Quantum dot-NanoLuc bioluminescence resonance energy transfer enables tumor imaging and lymph node mapping in vivo. Chem Commun(Camb)2016;52:6997-7000.
[16] Jiang TT, Yin NQ, Liu L, Song J, Huang QP, Zhu LX, et al. Au nanoflower@Si02@CdTe/CdS/ZnS quantumdot multi-functional nanoprobe for photothermal treatment and cellular imaging. RSC Adv 2014;4:23630-23636.
[17] Sugawa K, Tamura T, Tahara H, Yamaguchi D, Akiyama T, Otsuki J, et al. Metal-enhanced fluorescence platforms based on plasmonic ordered copper arrays:wavelength dependence of quenching and enhancement effects. ACS Nano 2013;7:9997-10010.
[18] Yuan L, Zhu J, Ren Y, Bai S. Sectional area-dependent plasmonic shifting in the truncated process of silver nanoparticles:from cube to octahedron. J Nanopart Res 2011;13:6305-6312.
[19] Sabaeian M, Khaledi-Nasab A. Size-dependent intersubband optical properties of dome-shaped InAs/GaAs quantum dots with wetting layer. Appl Opt2012;51:4176-4185.
[20] Naumova AV, Modo M, Moore A, Murry CE, Frank JA. Clinical imaging in regenerative medicine. Nat Biotechnol 2014;32:804-818.
[21] Hong GS, Antaris AL, Dai HJ. Near-infrared fluorophores for biomedical imaging. Nature Bio Eng 2017;1:0010.
[22] Hyun H, Owens EA, Wada H, Levitz A, Park G, Park MH, et al. Cartilage-specific near-infrared fluorophores for biomedical imaging. Angew Chem Int Ed Engl2015;54:8648-8652.
[23] Legant WR, Shao L, Grimm JB, Brown TA, Milkie DE, Avants BB, et al. High--density three-dimensional localization microscopy across large volumes. Nat Methods 2016;13:359-365.
[24] Cui W, Li J, Zhang Y, Rong H, Lu W, Jiang L. Effects of aggregation and the surface properties of gold nanoparticles on cytotoxicity and cell growth.Nanomedicine 2012;8:46-53.
[25] Kolny-Olesiak J, Weller H. Synthesis and application of colloidal CuInS2 semiconductor nanocrystals. ACS Appl Mater Interfaces 2013;5:12221-12237.
[26] Chen Y, Li S, Huang L, Pan D. Low-cost and gram-scale synthesis of watersoluble Cu-In-S/ZnS core/shell quantum dots in an electric pressure cooker.Nanoscale 2014;6:1295-1298.
[27] Shen S,Zhao L, Zhou Z, Guo L. Enhanced photocatalytic hydrogen evolution over Cu-doped ZnIn2S4 under visible light irradiation. J Phys Chem C