Abstract
Urinary proteins are an important component in urine samples that can be used as biological indicators for clinical diagnosis of diseases, such as proteinuria, kidney failure, diseases in the bladder, urinary tract and related organs. In this present work, we investigated candidate protein biomarkers in urine for diagnosis of cervical cancer using proteomic approach. The urine samples were concentrated and de-salted through ultrafiltration (3 kDa), followed by Spin-50 mini-sephadex column and were then separated by two-dimensional electrophoresis (2-DE) and annotated by mass spectrometer. Comparing 2-DE results between individual healthy donors and cervical cancer, patients showed that 4 urinary proteins; Protocadherin-8, Aryl hydrocarbon receptor nuclear translocatorlike protein 2, Serum albumin and Endorepellin, C-terminal domain V of perlecan were overexpressed by two-fold in the patient samples. Only the endorepellin LG3 fragment was validated by Western blot analysis. We also used 2D Western blot and nano LC-MS/MS to confirm the urinary biomarker as endorepellin LG3 fragment. Our study suggests that proteomics approach may be useful for the study of urinary proteins and provides an opportunity to discovery aberrant protein-associated diseases. We also revealed the candidate biomarkers that may be applied for cervical cancer diagnosis which benefits for medical treatment.
Highlights
According to reports from the American Cancer Society (ACS) since 2007 (National Cancer Institute, Centers of Disease Control and Prevention, North American Association of Central Cancer Registries and National Center for Health Statistic), in the USA, estimated 1,638,910 new cancer cases and 577,190 cancer deaths were diagnosed in 2012
Urine contains body filtrates such as water, salts, electrolytes and nitrogenous waste products and trace amounts of protein originating from blood plasma, the kidneys and the urogenital tract [16,17]
We found that one tryptic peptide that matched to perlecan by MS/MS, as well as PMF, resided on the C-terminal domain V of perlecan, commonly known as endorepellin (Table 2)
Summary
According to reports from the American Cancer Society (ACS) since 2007 (National Cancer Institute, Centers of Disease Control and Prevention, North American Association of Central Cancer Registries and National Center for Health Statistic), in the USA, estimated 1,638,910 new cancer cases and 577,190 cancer deaths were diagnosed in 2012. Cervical cancer was the third most common type of cancer and was the fourth most frequent cause of cancer death in women worldwide, with 529,512 (8.8%) new cases and 274,967 (8.2%) cancer death in 2008 [2]. Infection with human papillomavirus (HPV) is established as an initiating cause of cervical cancer, cooperating with other co-factors necessary for progression of this cancer, such as tobacco smoking, high parity, long-term hormonal contraceptive use and co-infection with HIV. The death rate of cervical cancer is still high [10]. Many researchers tend to investigate biological indicators of cancer status and progression before clinical symptoms or enabling real-time monitoring of drug response
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