Gene Trapping Identifies Chloride Channel 4 As a Novel Inducer of Colon Cancer Cell Migration, Invasion and Metastases

Overview

Journal Br J Cancer

Specialty Oncology

Date 2010 Jan 21

PMID 20087350

Citations 6

Authors

T Ishiguro

H Avila

S-Y Lin

T Nakamura

M Yamamoto

D D Boyd

Affiliations

Soon will be listed here.

Abstract

Background: To date, there are few reports on gene products contributing to colon cancer progression.

Methods: We used a gene trap comprised of an enhanced retroviral mutagen (ERM) cassette that includes a tetracycline-responsive promoter upstream of a haemagglutinin (HA) tag and a splice donor site. Integration of the ERM within an endogenous gene yields a tetracycline-regulated HA-tagged transcript. We transduced RKO colon cancer cells expressing a tetracycline trans-activator-off with the ERM-encoding retrovirus and screened for enhanced migration.

Results: One clone showed fivefold enhanced migration with tetracycline withdrawal. Rapid amplification of cDNA ends identified the trapped gene as the chloride channel 4 (CLCN4) exchanger. Stable expression of a CLCN4 cDNA enhanced motility, whereas cells knocked down or null for this transcript showed reduced migration/invasion. CLCN4-overexpressing RKO colon cancer cells were more resistant than controls to proton load-induced cytotoxicity, consistent with the H(+)-extruding function of this antiporter. Intra-splenic delivery of RKO-CLCN4 transfectants, but not controls, yielded liver metastases, and transcript levels were higher in colon cancer metastases to the liver when compared with primary tumours.

Conclusions: CLCN4 is a novel driver of colon cancer progression.

Citing Articles

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