Preliminary Proteomic Analysis of Indomethacin's Effect on Tumor Transplanted with Colorectal Cancer Cell in Nude Mice

Abstract

Nonsteroidal anti-inflammatory drugs such as indomethacin (IN) can exert anti-colorectal cancer (CRC) activity through cyclooxygenase independent mechanism, but the exactly biological mechanism is not completely known. Here we use proteomic tools to investigate the molecular mechanism of this action. First, nude mice bearing tumors derived from subcutaneous injection with human CRC cell line HCT116 were randomly allocated to groups treated with or without indomethacin. Later, tumor lumps were incised and then total proteins extracted. After separated with two-dimensional electrophoresis, thirty-one differently expressed spots were found between IN-treated and non-IN-treated groups, of which 25 spots decreased and 6 spots increased in abundance in IN-treated group. Through matrix-assisted laser desorption ionization time of flight mass spectrometry and then NCBInr and SWISS-PROT databases searching, 12 protein spots were finally identified including galectin-1, annexin A1, annexin IV, transcription factor BTF3A, calreticulin. Most of the identified proteins are correlated with tumor's biological properties of proliferation, invasion, apoptosis and immunity, or take part in cell's signal transduction. From above we thought that indomethacin can exert its effect on colorectal cancer through regulating several proteins' expression directly or indirectly. Further study of these proteins may be helpful in founding new targets of drugs for cancer chemotherapy.

Improving Colorectal Cancer Management: The Potential of Proteomics

“…Tissue 2D-GE [145] Aldolase A + Tissue 2D-GE [146] Aldolase C, fructose-biphosphate + Tissue 2D-GE [37] Aldolase, fructose-biphosphate + Tissue 2D-GE [20] [ 36] Cell line 2D-DIGE -Cell line 2D-GE [36] NR Tissue 2D-GE [140] α-1 antitrypsin precursor…”

“…Tissue 2D-GE [22,142,147] 2D-DIGE α-3 type VI collagen, isoform 2 precursor + Tissue 2D-GE [146] α-1B glycoprotein -Tissue 2D-GE [143] α(B)-crystallin -Tissue 2D-GE [145] α-defensins (human neutrophil peptide) 1-3 + Tissue SELDI-TOF MS [57][58][59] serum Annexin I (A1) + Tissue 2D-GE [37] [142] 2D-DIGE Annexin A2 (Annexin II) + Tissue 2D-GE [140,146] [ 141,148] Cell line…”

“…future science group future science group [41,45] Δ (3,5)-δ-(2,4)-dienoyl-CoA isomerase, mitochondrial + Tissue 2D-GE [37] Tissue 2D-GE [139] 5-lipoxygenase + Tissue 2D-GE [41,45] 11s regulator chain B NR Tissue 2D-GE [140] Cell line 2D-GE [141] 14-3-3 β/α protein + Tissue 2D-DIGE [142] 14-3-3 β + Tissue 2D-GE [22] 14-3-3 γ protein + Tissue 2D-DIGE [142] 14-3-3 ε -Tissue 2D-GE [22] 14-3-3 protein ζ/δ (protein kinase C inhibitor protein 1) + Tissue 2D-DIGE [142] -(*) Tissue 2D-GE [22] [ 33] Cell line [41,45] 14-3-3 θ + Tissue 2D-GE [25] -(*) Tissue 2D-GE [43] 60S acidic ribosomal protein P0 (L10E) + Cell line 2D-GE [23] Aconitase 2 -Tissue 2D-GE [20] Aconitate hydratase, mitochondrial precursor -Tissue 2D-GE [20] Actin fragment + Tissue 2D-DIGE [142] Tissue 2D-DIGE [142] Actin α -Tissue 2D-GE [22,143] Actin β + (*) Tissue 2D-GE [22] -(*) Tissue 2D-GE [41,43,45,143] NR Tissue 2D-GE [140] [ 144] Actin γ -(*) Tissue 2D-GE [41,43,45] Activator of 90 Tissue 2D-GE [145] Adenosyl homocysteinase and eukaryotic initiation factor 4A like + Tissue 2D-DIGE [142] Adenylyl cyclase-associated protein + Tissue 2D-GE [146] Albumin -Tissue 2D-GE …”

“…Some have investigated cellular pathways involved in the development of CRC [23][24][25][26][27], others assessed proteins implicated in the development of metastases [28][29][30]. In addition, protein profiles reflecting the sensitivity of CRC cells to therapeutic agents [31][32][33] or the effect of diet [34,35] and therapeutic future science group future science group www.futuremedicine.com compounds on colorectal cells [36,37] have been the subject of preclinical experiments. However, discrepancies between cells grown in vitro and in vivo exist (e.g., because of adaptation to cell culture conditions [38], as well as differences between animals and humans [39]).…”

Keratin 17 identified by proteomic analysis may be involved in tumor angiogenesis

“…Candidate protein spots were cut off from the gels and extracted by in gel digestion as described by Wang et al (27). The generated peptides were analyzed by MALDI-TOF-MS us-http://bmbreports.org ing a Bruker Daltonics AutoFlex TOF-TOF LIFT mass spectrometer (Germany).…”