Combination of CALR and PDIA3 is a potential prognostic biomarker for non-small cell lung cancer

Abstract

Proteomic-based approaches for biomarker discovery are promising strategies used in cancer research. In this study, we performed quantitative proteomic analysis on 16 paired samples of non-small cell lung cancer (NSCLC) and adjacent non-tumor lung tissues using label-free quantitative proteomics and liquid chromatography-tandem mass spectrometry/mass spectrometry (LC-MS/MS) to identify differentially expressed proteins. A total of 91 proteins were differentially expressed in NSCLC compared with adjacent non-tumor lung tissues among 4047 identified proteins (fold change > 1.5 or < 0.67, P < 0.05). Gene ontology (GO) analysis, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and ingenuity pathway analysis (IPA) of 91 dysregulated proteins showed that they were related to the cancer-associated biological processes. We confirmed that the candidate proteins, calreticulin (CALR) and protein disulfide isomerase family A member 3 (PDIA3) were overexpressed in NSCLC by real-time PCR using 20 paired samples and western blot using 5 paired samples. PDIA3 expression was highly associated with CALR expression (Spearman r = 0.345, P = 0.001) and they were co-localized and interacted with each other in A549 and H460 cells. Moreover, survival analysis performed in tissue microarray with 88 samples indicated that low expression of both CALR and PDIA3 in NSCLC was positively associated with poor overall survival. Combination of CALR and PDIA3 might serve as an efficient biomarker and improved the prediction of NSCLC prognosis significantly (P = 0.023). Our results collectively provide a potential biomarker dataset for NSCLC prognosis, especially the prognostic value of combined expression of CALR and PDIA3.

Calreticulin Blockade Attenuates Murine Acute Lung Injury by Inducing Polarization of M2 Subtype Macrophages

“…A body of evidences showed that CALR is highly expressed in tumor cells and transported onto cell membrane to serve as a C1q receptor and prophagocytic signal for macrophage phagocytosis of cancer cells and dead cells (12,13). Because the exposed CALR can be released into extracellular milieu (14,15), the serum CALR is highly elevated in the tumor patients and positively correlated to the prognosis of tumor (16,17). Therefore, serum CALR is considered as a useful biomarker in cancer diagnosis and prognosis evaluation.…”

Identification of differentially expressed genes in non-small cell lung cancer

“…Twenty paired samples were excluded from analysis owing to incomplete patient information and/or sample absence. Therefore, IHC was performed as reported previously [4] on 70 matched specimens. Paraffin sections were dewaxed, followed by antigen retrieval with Tris-EDTA buffer (pH 9).…”

“…Hematoxylin was used to counterstain the nuclei. Intensity and density of TOP2A-positive cells was evaluated and scored as reported before [4].…”

“…While some success is being achieved with newer immunological and targeted therapies for NSCLC, there are still significant limitations precluding their use in many cases [3]. Notwithstanding, the low 5-year survival rate for patients with lung cancer is largely due to insufficient preventive efforts and generalized late diagnosis [4].…”

Contribuições para o estudo da heterogeneidade inter e intratumoral em neoplasias mamárias em cadelas

“…In addition, for canine mammary cancer, immunostaining for Derlin-1 showed that metastatic cells, the later stage of tumor progression, presented an increased stress-associated unfolded protein response (KLOPFLEISCH et al, 2010a) compared to non-metastatic samples.PDIA3 interacts with lectin chaperones calreticulin and calnexin to modulate folding of newly synthesized glycoproteinsBAIG;WILLIAMS, 2006). The low expression of PDIA3 have been associated to a poor overall survival for non-small lung cancer and gastric cancer, due to the formation of a complex with MHC class I and the high correlation with the expression of calreticulin, responsible for maintaining adequate calcium levels in organisms and also acting in ERBAIG;WILLIAMS, 2006;WANG et al, 2017b;SHIMODA et al, 2019). Calnexin is a highly abundant transmembrane chaperone which folds synthesized glycoproteins in the lumen of the endoplasmic reticulum(AEBI et al, 2010;SHIBATA et al, 2010), while the heat shock protein 5 (also known as BIP) is a master regulator of the UPR (MALHOTRA;KAUFMAN, 2007).…”