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Intracellular Acidosis Promotes Mitochondrial Apoptosis Pathway: Role of EMMPRIN Down-regulation via Specific Single-chain Fv Intrabody
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Document Title
Intracellular Acidosis Promotes Mitochondrial Apoptosis Pathway: Role of EMMPRIN Down-regulation via Specific Single-chain Fv Intrabody
Author
Thammasit P, Sangboonruang S, Suwanpairoj S, Khamaikawin W, Intasai N, Kasinrerk W, Tayapiwatana C, Tragoolpua K
Name from Authors Collection
Affiliations
Chiang Mai University; Chiang Mai University; Chiang Mai University; Chiang Mai University; National Science & Technology Development Agency - Thailand; Chiang Mai University
Type
Article
Source Title
JOURNAL OF CANCER
ISSN
1837-9664
Year
2015
Volume
6
Issue
2-3
Open Access
Green Published, gold, Green Submitted
Publisher
IVYSPRING INT PUBL
DOI
10.7150/jca.10879
Format
Abstract
Extracellular matrix metalloproteinase inducer (EMMPRIN) is a human leukocyte surface molecule that is enriched on the surface of many cancer cells, and it plays an important role in proliferation and metastasis. In this study, we utilized the chimeric adenoviral vector Ad5/ F35 carrying gene encoding scFv against EMMPRIN (scFv-M6-1B9) to down-regulate EMMPRIN cell surface expression and investigated programmed cell death response in colorectal cancer (CRC) cell, Caco-2. The scFv-M6-1B9 intrabody exhibits robust activity in reducing EMMPRIN cell surface expression. This approach led to the inducing of apoptosis, which was relative to the increasing of apoptotic bodies in sub-G1 peak, phosphatidylserine externalization, as well as TUNEL-positive cells. In addition, real-time RT-PCR and western blotting analysis indicated that apoptosis was enhanced through the mitochondrial pathway, a marked reduction of Bcl-2, leading to the translocation of cytochrome c and also the dramatic activation of caspase-3. Moreover, carcinoembryonic antigen (CEA), a tumor marker for CRC, was found to have significantly diminished in both secreted protein and mRNA levels. In conclusion, these findings suggest that EMMPRIN down-regulation by scFv-M6-1B9 intrabody has great potential in enhancing the efficacy of apoptosis induction through the mitochondrial pathway and in effecting a decline in the CEA level. Thus, its benefits could be applied to project the future prospects for targeted gene therapy and therapeutic application in monitoring colorectal cancer.
Funding Sponsor
Discovery-based Development Grant; Research Chair Grant from the National Sciences and Technology Development Agency (NSTDA); Thailand Research Fund (TRF); National Research University Project under the Thailand Office of the Commission on Higher Education, Thailand
License
CC BY-NC
Rights
Publisher
Publication Source
WOS