Nonsteroidal anti-inflammatory drug sulindac sulfide suppresses structural protein Nesprin-2 expression in colorectal cancer cells

• Published in: Biochimica et biophysica acta 2014-01, Vol.1840 (1), p.322-331
• Main Authors: Liggett, Jason L., Choi, Chang Kyoung, Donnell, Robert L., Kihm, Kenneth D., Kim, Jong-Sik, Min, Kyung-Won, Noegel, Angelika Anna, Baek, Seung Joon
• Format: Article
• Language: English
• Subjects: Anti-Inflammatory Agents, Non-Steroidal - pharmacology; Biomarkers, Tumor - genetics; Biomarkers, Tumor - metabolism; Blotting, Western; Cell Adhesion - drug effects; Cell Proliferation - drug effects; Colon cancer; Colorectal Neoplasms - drug therapy; Colorectal Neoplasms - metabolism; Colorectal Neoplasms - pathology; Electric Impedance; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic - drug effects; Humans; Immunoenzyme Techniques; Male; Micro-impedance; Microfilament Proteins - antagonists & inhibitors; Microfilament Proteins - genetics; Microfilament Proteins - metabolism; NAG-1; Nerve Tissue Proteins - antagonists & inhibitors; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Nesprin-2; NSAID; NUANCE; Nuclear Proteins - antagonists & inhibitors; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Oligonucleotide Array Sequence Analysis; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA Stability - drug effects; RNA, Messenger - genetics; RNA, Small Interfering - genetics; Sulindac - analogs & derivatives; Sulindac - pharmacology; Tissue Array Analysis; Tumor Cells, Cultured
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2013.09.032

Nonsteroidal anti-inflammatory drugs (NSAIDs) are well known for treating inflammatory disease and have been reported to have anti-tumorigenic effects. Their mechanisms are not fully understood, but both cyclooxygenase (COX) dependent and independent pathways are involved. Our goal was to shed further light on COX-independent activity. Human colorectal cancer cells were observed under differential interference contrast microscopy (DICM), fluorescent microscopy, and micro-impedance measurement. Microarray analysis was performed using HCT-116 cells treated with sulindac sulfide (SS). PCR and Western blots were performed to confirm the microarray data and immunohistochemistry was performed to screen for Nesprin-2 expression. Micro-impedance was repeating including Nesprin-2 knock-down by siRNA. HCT-116 cells treated with SS showed dramatic morphological changes under DICM and fluorescent microscopy, as well as weakened cellular adhesion as measured by micro-impedance. Nesprin-2 was selected from two independent microarrays, based on its novelty in relation to cancer and its role in cell organization. SS diminished Nesprin-2 mRNA expression as assessed by reverse transcriptase and real time PCR. Various other NSAIDs were also tested and demonstrated that inhibition of Nesprin-2 mRNA was not unique to SS. Additionally, immunohistochemistry showed higher levels of Nesprin-2 in many tumors in comparison with normal tissues. Further micro-impedance experiments on cells with reduced Nesprin-2 expression showed a proportional loss of cellular adhesion. Nesprin-2 is down-regulated by NSAIDs and highly expressed in many cancers. Our data suggest that Nesprin-2 may be a potential novel oncogene in human cancer cells and NSAIDs could decrease its expression. •NSAIDs affect cell–cell interaction and membrane permeability.•Nesprin-2 is likely expressed in tumors.•Nesprin-2 may possess possible oncogenic activity.•NSAIDs decrease the expression of Nesprin-2.