Identification of Differentially-Expressed Proteins between Early Submucosal Non-Invasive and Invasive Colorectal Cancer Using 2D-DIGE and Mass Spectrometry

Abstract

Early detection and diagnosis of colorectal cancer (CRC) are closely related to a better therapeutic outcome, and the five-year survival rate ofearly CRC is over 90%. Though endoscopic minimally invasive treatment has become a quick and effective therapy for early CRC, endoscopic biopsies are usually not deep enough to obtain tissues from the submucosal layer and it is difficult to determine whether early CRC has infiltrated into the submucosa. Therefore, in the present study, we constructed tumor models of early submucosal non-invasive CRC (SNICRC) and submucosal invasive CRC (SICRC) in Fischer-344 rats induced by N-methyl-N-nitrosourea (MNU). The differentially-expressed proteins were analyzed and identified in SNICRC, SICRC and normal control (NC) tissues using highly sensitive twodimensional differential gel electrophoresis (2D-DIGE) coupled with mass spectrometry (MS). Proteomic data revealed 132 protein spots between SNICRC and SICRC, 162 protein spots between SICRC and NC and 154 protein spots between SNICRC and NC which were found differentially expressed. These differential spots were picked, in-gel digested and peptide mass fingerprints were obtained by MALDI-TOF-MS/MS. Finally, five differentially-expressed proteins in SNICRC, SICRC and NC were identified, and increases in Transgelin, peptidylprolyl isomerase A (PPIA) and tropomyosin alpha isoform d were observed, while decreases in carbonic anhydrase 2 (CAlI) and an unnamed protein were detected in SICRC compared with SNICRC and NC. Furthermore, Fluorescence-based quantitative polymerase chain reaction (FQ-PCR), Western blotting and immunohistochemistry assays also revealed significant up regulation of Transgelin expression and down-regulation of CAlI expression in SICRC tissues. In conclusion,~D-DIGE is confirmed to be an efficient strategy that enables us to identify differentiallyexpressed proteins between early SNICRC and SICRC. The potential biomarkers such as Transgelin and CAlI may be used for the detection of early SICRC Colorectal cancer (CRC) is a major cause of cancer incidence and mortalit y, and there are nearly one million new cases of CRC diagnosed worldwide each year and half a million deaths (1). The past two decades have brought major advances in CRC screening and therapeuti cs including advances in colonoscop y therapy. However, it is hard for endoscopic examination or other means to detect early CRC lesions.Thus, research focuses on specific strategies

Transgelin is a TGFβ-inducible gene that regulates osteoblastic and adipogenic differentiation of human skeletal stem cells through actin cytoskeleston organization

“…TAGLN has previously been implicated in cell migration; 6, 15, 16, 17, 18, 19 however, the role of TAGLN expression in hMSC migration has not been studied. Thus, we studied the effects of TAGLN deficiency and overexpression on hMSC cell migration in two independent assays.…”

Transgelin Silencing Induces Different Processes in Different Breast Cancer Cell Lines

“…However, TAGLN has both positive and negative roles in these processes, depending on the cell line. [ 10,17–20 ]…”

Mass Spectrometry-Based Proteomics in Molecular Diagnostics: Discovery of Cancer Biomarkers Using Tissue Culture

“…This technique is routinely used for the discovery of candidate biomarkers as well as any quantitative proteomic data generation and therapeutic drug development. Zhang et al used this technique for the identification of differentially expressed proteins between early submucosal noninvasive and invasive colorectal cancer [37]. They have established a Fischer-344 rat model for the invasive and noninvasive colorectal cancer and found two candidates, transgelin (upregulated) and carbonic anhydrase 2 (CSII) to play significant role in CRC.…”

“…They have established a Fischer-344 rat model for the invasive and noninvasive colorectal cancer and found two candidates, transgelin (upregulated) and carbonic anhydrase 2 (CSII) to play significant role in CRC. They have also validated these candidates through fluorescence-based quantitative polymerase chain reaction, western blotting and immunohistochemistry assays [37]. In a similar kind of study, isocitrate dehydrogenase 1 (IDH1) was detected and validated as a potential biomarker for nonsmall cell lung carcinoma [38].…”