Proteomics Approaches for Identification of Tumor Relevant Protein Targets in Pulmonary Squamous Cell Carcinoma by 2D-DIGE-MS

Abstract

Potential markers for progression of pulmonary squamous cell carcinoma (SCC) were identified by examining samples of lung SCC and adjacent normal tissues using a combination of fluorescence two-dimensional difference gel electrophoresis (2D-DIGE), matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and electrospray ionization quadrupole-time of flight mass spectrometry (ESI-Q-TOF). The PANTHER System was used for gel image based quantification and statistical analysis. An analysis of proteomic data revealed that 323 protein spots showed significantly different levels of expression (P≤0.05) in lung SCC tissue compared to expression in normal lung tissue. A further analysis of these protein spots by MALDI-TOF-MS identified 81 different proteins. A systems biology approach was used to map these proteins to major pathways involved in numerous cellular processes, including localization, transport, cellular component organization, apoptosis, and reproduction. Additionally, the expression of several proteins in lung SCC and normal tissues was examined using immunohistochemistry and western blot. The functions of individual proteins are being further investigated and validated, and the results might provide new insights into the mechanism of lung SCC progression, potentially leading to the design of novel diagnostic and therapeutic strategies.

Quantitative Proteomic Profiling the Molecular Signatures of Annexin A5 in Lung Squamous Carcinoma Cells

“…According to the results from TCGA, the expression levels of ANXA5 in NSCLC tissues were significantly lower in lung squamous cell carcinoma (LUSC) and lung adenocarcinoma (LUAD) ( Table 1 ) compared to matched normal samples, which is consistent with our proteomic result from LUSC and adjacent normal tissues [ 6 , 21 ].…”

“…In our previous report [ 6 , 21 ], ANXA5 was found to be down-regulated in lung squamous cell carcinoma (LUSC), compared with adjacent normal tissues, which is consistent with our result conducted from The Cancer Genome Atlas (TCGA). We have demonstrated that upregulation of ANXA5 inhibits the proliferation, migration, and invasion abilities of NCI-H520 cells in vitro.…”

“…Details about these 70 proteins, including protein ID, protein name, gene name and fold change regulated by ANXA5, are presented in Table 2 . It is worth noting that, among these identified proteins, five proteins (Hsp90, ENO1, ALDOA, PDIA6, HSPA5) were reported as the differential proteins identified in lung cancer tissues as compared to normal lung in our previous work [ 21 ].…”

Elevated eukaryotic elongation factor 2 expression is involved in proliferation and invasion of lung squamous cell carcinoma

“…These proteins could serve as early diagnosis molecular markers or prognostic indicators. In the previous research, we compared the protein profiles between LSCC tissues and adjacent normal lung tissues, and we identified a number of differentially expressed proteins by using 2D-DIGE and MS approaches [ 3 ]. Among these protein candidates, we were particularly interested in the eukaryotic elongation factor 2 (EF2), a key enzyme in the elongation step in protein translation.…”

Advances of Salivary Proteomics in Oral Squamous Cell Carcinoma (OSCC) Detection: An Update

“…The high morbidity and mortality rate in OSCC patients [5] ultimately led to the aspiration for enhanced knowledge of the characteristics and pathogenesis involved, as clinical and histological analyses are the only basis for OSCC diagnosis, furthermore, specific biomarkers are highly supportive of untraceable hidden sites of OSCC and for the screening of high risk patients [4]. Since OSCC is a multifactorial disease, molecular pathways are necessary for diagnostics, prognostics and treatment of cancer [2], for which the human genome database (HGD), the proteomic approach, introduced by novel technology, is developed to monitor genetic alterations and to recognize protein biomarkers linked with growth, progression and recurrence of the tumour [6]. In comparison to tissue biopsy which is invasive, there have been plenty of biomarkers identified, amongst them, the most recommended biomarker detection medium for OSCC includes biomarkers in body fluids, such as blood and saliva (serum and plasma) [7].…”