Quantitative electrochemical detection of cathepsin B activity in breast cancer cell lysates using carbon nanofiber nanoelectrode arrays toward identification of cancer formation.

UNLABELLED The proteolytic activity of cathepsin B in complex breast cell lysates has been measured with alternating current voltammetry (ACV) using ferrocene (Fc)-labeled-tetrapeptides immobilized on nanoelectrode arrays (NEAs) fabricated with vertically aligned carbon nanofibers (VACNFs). Four types of breast cells have been tested, including normal breast cells (HMEC), transformed breast cells (MCF-10A), breast cancer cells (T47D), and metastatic breast cancer cells (MDA-MB-231). The detected protease activity was found increased in cancer cells, with the MDA-MB-231 metastatic cancer cell lysate showing the highest cathepsin B activity. The equivalent cathepsin B concentration in MDA-MB-231 cancer cell lysate was quantitatively determined by spiking recombinant cathepsin B into the immunoprecipitated MDA-MB-231 lysate and the HMEC whole cell lysate. The results illustrated the potential of this technique as a portable multiplex electronic device for cancer diagnosis and treatment monitoring through rapid profiling the activity of specific cancer-relevant proteases. FROM THE CLINICAL EDITOR Breast cancer is the most common cancer in women. In this report, the authors applied the technique of nanoelectrode arrays to try to detect and compare cathepsin B activities in normal and breast cancer cells. It was found that protease activity correlated positively with the degree of malignancy cancer cells. Taking this further, this technique may be useful for rapid diagnosis of cancer in the future.

[1]  H. Muss Endocrine therapy for advanced breast cancer: A review , 2005, Breast Cancer Research and Treatment.

[2]  Yun Zheng,et al.  Detection of cathepsin B, cathepsin L, cystatin C, urokinase plasminogen activator and urokinase plasminogen activator receptor in the sera of lung cancer patients. , 2011, Oncology letters.

[3]  Jun Li,et al.  Characterization of carbon nanofiber electrode arrays using electrochemical impedance spectroscopy: effect of scaling down electrode size. , 2010, ACS nano.

[4]  J. Mort Chapter 406 – Cathepsin B , 2013 .

[5]  Alan J. Barrett,et al.  [41] Cathepsin B, cathepsin H, and cathepsin L , 1981 .

[6]  M. King Breast cancer genes: how many, where and who are they? , 1992, Nature Genetics.

[7]  Bonnie F. Sloane,et al.  The cysteine protease cathepsin B in cancer , 1996 .

[8]  M. Morrow,et al.  Molecular mechanisms of resistance to tamoxifen therapy in breast cancer. , 1993, Archives of surgery.

[9]  Bonnie F. Sloane,et al.  Increased gelatinase A (MMP-2) and cathepsin B activity in invasive tumor regions of human colon cancer samples. , 1994, The American journal of pathology.

[10]  K. Inoue,et al.  Immunohistochemical study of cathepsin B. Prognostic significance in human lung cancer , 1994, Cancer.

[11]  L. Juliano,et al.  Positional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides to define substrate specificity of carboxydipeptidases: assays with human cathepsin B. , 2004, Analytical biochemistry.

[12]  M. Babjuk,et al.  Urine and Serum Cathepsin B Concentrations in the Transitional Cell Carcinoma of the Bladder , 2012, Journal of clinical laboratory analysis.

[13]  E. Lissi,et al.  A theoretical approach to some analytical properties of heterogeneous enzymatic assays. , 2004, Analytical chemistry.

[14]  Bonnie F. Sloane,et al.  Unraveling the role of proteases in cancer. , 2000, Clinica chimica acta; international journal of clinical chemistry.

[15]  V. Jordan Alternate antiestrogens and approaches to the prevention of breast cancer , 1995, Journal of cellular biochemistry. Supplement.

[16]  C. Redmond,et al.  Endocrine treatment of breast cancer in women. , 1990, Endocrine reviews.

[17]  E. Lam,et al.  Electron Transfer at Electrodes through Conjugated “Molecular Wire” Bridges , 1999 .

[18]  Lateef U. Syed,et al.  Electrochemical Protease Biosensor Based on Enhanced AC Voltammetry Using Carbon Nanofiber Nanoelectrode Arrays. , 2013, The journal of physical chemistry. C, Nanomaterials and interfaces.

[19]  Jun Li,et al.  Inlaid Multi-Walled Carbon Nanotube Nanoelectrode Arrays for Electroanalysis , 2005 .

[20]  M. Meyyappan,et al.  Carbon Nanotube Nanoelectrode Array for Ultrasensitive DNA Detection , 2003 .

[21]  D. Hua,et al.  Quantitative electrochemical detection of cathepsin B activity in complex tissue lysates using enhanced AC voltammetry at carbon nanofiber nanoelectrode arrays. , 2014, Biosensors & bioelectronics.

[22]  Neil D. Rawlings,et al.  Handbook of proteolytic enzymes , 1998 .

[23]  D. Hua,et al.  Inhibition of norovirus 3CL protease by bisulfite adducts of transition state inhibitors. , 2013, Bioorganic & medicinal chemistry letters.

[24]  Jun Li,et al.  Wafer-scale fabrication of patterned carbon nanofiber nanoelectrode arrays: a route for development of multiplexed, ultrasensitive disposable biosensors. , 2009, Biosensors & bioelectronics.

[25]  Bonnie F. Sloane,et al.  Cathepsin B: a potential prognostic marker for inflammatory breast cancer , 2011, Journal of Translational Medicine.

[26]  Lateef U. Syed,et al.  Enhanced Electron Transfer Rates by AC Voltammetry for Ferrocenes Attached to the End of Embedded Carbon Nanofiber Nanoelectrode Arrays. , 2011, Electroanalysis.

[27]  V. Jordan,et al.  Development of antiestrogens and their use in breast cancer: eighth Cain memorial award lecture. , 1990, Cancer research.

[28]  S. Ethier Growth factor synthesis and human breast cancer progression. , 1995, Journal of the National Cancer Institute.

[29]  S. Kostrov,et al.  Handbook of proteolytic enzymes, 3rd Edition , 2013 .

[30]  S. Hilsenbeck,et al.  Prognostic factors: Rationale and methods of analysis and integration , 2004, Breast Cancer Research and Treatment.

[31]  Erik Sahai,et al.  Mechanisms of cancer cell invasion. , 2005, Current opinion in genetics & development.

[32]  R. Fridman,et al.  Extracellular proteases as targets for treatment of cancer metastases. , 2004, Chemical Society reviews.

[33]  R. Dickson,et al.  Growth factors in breast cancer. , 1995, Endocrine reviews.