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

Abstract

BACKGROUND:

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.

METHODS:

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.

RESULTS:

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.

CONCLUSIONS:

Nesprin-2 is down-regulated by NSAIDs and highly expressed in many cancers.

GENERAL SIGNIFICANCE:

Our data suggest that Nesprin-2 may be a potential novel oncogene in human cancer cells and NSAIDs could decrease its expression.

• Authors: Jason L Liggett, Chang Kyoung Choi, Robert L Donnell, Kenneth D Kihm, Jong-Sik Kim, Kyung-Won Min, Angelika Anna Noegel, Seung Joon Baek
• Journal: Biochimica et biophysica acta (Biochim Biophys Acta) Vol. 1840 Issue 1 Pg. 322-31 (Jan 2014) ISSN: 0006-3002 [Print] Netherlands
• PMID: 24080406 (Publication Type: Journal Article)
• Copyright: © 2013.

Chemical References

• Anti-Inflammatory Agents, Non-Steroidal
• Biomarkers, Tumor
• Microfilament Proteins
• Nerve Tissue Proteins
• Nuclear Proteins
• RNA, Messenger
• RNA, Small Interfering
• SYNE2 protein, human
• Sulindac
• sulindac sulfide

Topics

• Anti-Inflammatory Agents, Non-Steroidal (pharmacology)
• Biomarkers, Tumor (genetics, metabolism)
• Blotting, Western
• Cell Adhesion (drug effects)
• Cell Proliferation (drug effects)
• Colorectal Neoplasms (drug therapy, metabolism, pathology)
• Electric Impedance
• Female
• Gene Expression Profiling
• Gene Expression Regulation, Neoplastic (drug effects)
• Humans
• Immunoenzyme Techniques
• Male
• Microfilament Proteins (antagonists & inhibitors, genetics, metabolism)
• Nerve Tissue Proteins (antagonists & inhibitors, genetics, metabolism)
• Nuclear Proteins (antagonists & inhibitors, genetics, metabolism)
• Oligonucleotide Array Sequence Analysis
• RNA Stability (drug effects)
• RNA, Messenger (genetics)
• RNA, Small Interfering (genetics)
• Real-Time Polymerase Chain Reaction
• Reverse Transcriptase Polymerase Chain Reaction
• Sulindac (analogs & derivatives, pharmacology)
• Tissue Array Analysis
• Tumor Cells, Cultured