Signal amplification cytosensor for evaluation of drug-induced cancer cell apoptosis.

Apoptosis is involved in the pathology of a variety of diseases. The measurement of apoptosis will help us to evaluate the onset of disease and the effect of therapeutic interventions. In addition, the increased demand for understanding the early stages of apoptosis is pushing the envelope for solutions in early instance real-time monitoring of death kinetics. Here we present a novel electrochemiluminescent cytosensing strategy to quantitate apoptotic cell numbers, screen some anticancer drugs, and evaluate their effects on hepatocarcinoma cell line (HepG2) cells by utilizing the human antiphosphatidyl serine antibody (APSA) conjugated Ru(bpy)(3)(2+)-encapsulated silica nanoparticle (APSA-SiO(2)@Ru) as the detection probe. HepG2 cells were easily immobilized on the arginine-glycine-aspartic acid-serine (RGDS)-multiwalled carbon nanotubes (RGDS-MWCNTs) nanocomposite by the specific combination of RGD domains with integrin receptors on the cell surface. Then APSA-SiO(2)@Ru was introduced to the surface of apoptosis cells through the specific interaction between APSA and phosphatidylserine (PS) that distributed on the outer membrane of apoptotic cells. On the basis of the signal amplification of the APSA-SiO(2)@Ru nanoprobe, the cytosensor could respond as low as 800 cells mL(-1), showing very high sensitivity. In addition, the dynamic alterations of surface PS expression on HepG2 cells in response to drugs and the cell heterogeneity were also demonstrated. The strategy presented a promising platform for highly sensitive cytosensing and convenient screening of some clinically available anticancer drugs.

[1]  Luciano Bachmann,et al.  Application of FTIR Spectroscopy for Identification of Blood and Leukemia Biomarkers: A Review over the Past 15 Years , 2011 .

[2]  Albert van den Berg,et al.  Quantum dots based probes conjugated to annexin V for photostable apoptosis detection and imaging. , 2006, Nano letters.

[3]  Regina Luttge,et al.  Apoptotic cell death dynamics of HL60 cells studied using a microfluidic cell trap device. , 2005, Lab on a chip.

[4]  C. Creutz The annexins and exocytosis. , 1992, Science.

[5]  Wei Zhu,et al.  Detection of apoptosis based on the interaction between annexin V and phosphatidylserine. , 2009, Analytical chemistry.

[6]  G M Cohen,et al.  Caspases: the executioners of apoptosis. , 1997, The Biochemical journal.

[7]  D. Green,et al.  Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl , 1995, The Journal of experimental medicine.

[8]  H. Hug,et al.  Rhodamine 110-linked amino acids and peptides as substrates to measure caspase activity upon apoptosis induction in intact cells. , 1999, Biochemistry.

[9]  T. Cotter,et al.  Regulation and measurement of oxidative stress in apoptosis. , 2002, Journal of immunological methods.

[10]  V. Fadok,et al.  Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. , 1992, Journal of immunology.

[11]  Yafeng Wu,et al.  A novel electrochemiluminescence immunosensor via polymerization-assisted amplification. , 2010, Chemical communications.

[12]  Hsin-Yi Peng,et al.  Isolation of substances with antiproliferative and apoptosis-inducing activities against leukemia cells from the leaves of Zanthoxylum ailanthoides Sieb. & Zucc. , 2011, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[13]  Jun Liu,et al.  Analysis of nonadherent apoptotic cells by a quantum dots probe in a microfluidic device for drug screening. , 2009, Analytical chemistry.

[14]  C. Stroh,et al.  Death by a thousand cuts: an ever increasing list of caspase substrates , 1998, Cell Death and Differentiation.

[15]  H. Gratzner,et al.  Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: A new reagent for detection of DNA replication. , 1982, Science.

[16]  V. Fadok,et al.  The role of phosphatidylserine in recognition of apoptotic cells by phagocytes , 1998, Cell Death and Differentiation.

[17]  Guoli Shen,et al.  An Annexin V-based biosensor for quantitatively detecting early apoptotic cells. , 2009, Biosensors & bioelectronics.

[18]  Fei Deng,et al.  Elucidation of the reinforcing mechanism in carbon nanotube/rubber nanocomposites. , 2011, ACS nano.

[19]  Yafeng Wu,et al.  Enzyme-functionalized silica nanoparticles as sensitive labels in biosensing. , 2009, Analytical chemistry.

[20]  Joseph D. Gong,et al.  Carbon nanotube amplification strategies for highly sensitive immunodetection of cancer biomarkers. , 2006, Journal of the American Chemical Society.

[21]  Huangxian Ju,et al.  Lectin-based nanoprobes functionalized with enzyme for highly sensitive electrochemical monitoring of dynamic carbohydrate expression on living cells. , 2010, Analytical chemistry.

[22]  Chunlei Zhu,et al.  A potent fluorescent probe for the detection of cell apoptosis. , 2011, Chemical communications.

[23]  Songqin Liu,et al.  Electrochemiluminescence immunosensor for ultrasensitive detection of biomarker using Ru(bpy)(3)(2+)-encapsulated silica nanosphere labels. , 2010, Analytica chimica acta.

[24]  C. Reutelingsperger,et al.  Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure. , 1998, Cytometry.

[25]  P. Fraker,et al.  Versatility of merocyanine 540 for the flow cytometric detection of apoptosis in human and murine cells. , 2002, Journal of immunological methods.

[26]  W. Lu,et al.  Annexin A5–Conjugated Polymeric Micelles for Dual SPECT and Optical Detection of Apoptosis , 2011, The Journal of Nuclear Medicine.

[27]  Lei Liu,et al.  Electrochemical approach to detect apoptosis. , 2008, Analytical chemistry.

[28]  Soma Mukherjee,et al.  Spectroscopic, cytotoxic and DFT studies of a luminescent palladium(II) complex of a hydrazone ligand that induces apoptosis in human prostate cancer cells , 2011 .

[29]  Yanbin Li,et al.  Immunobiosensor chips for detection of Escherichia coil O157:H7 using electrochemical impedance spectroscopy. , 2002, Analytical chemistry.

[30]  T. Ravikumar,et al.  Apoptosis induced by cryo‐injury in human colorectal cancer cells is associated with mitochondrial dysfunction , 2003, International journal of cancer.

[31]  Y. Mély,et al.  Fluorescent biomembrane probe for ratiometric detection of apoptosis. , 2007, Journal of the American Chemical Society.

[32]  H. Lecoeur,et al.  Multiparametric flow cytometric analysis of biochemical and functional events associated with apoptosis and oncosis using the 7-aminoactinomycin D assay. , 2002, Journal of immunological methods.