Compensation of type I and type II cytokeratin pools in lung cancer

Abstract

Cytokeratins (CKs) constitute the largest family of intermediate filament proteins, and are subdivided into type I (CK9-CK23) and type II (CK1-CK8) subclasses. CK19 is expressed in non-small cell lung cancer (NSCLC), and serum CK19 fragment (referred to as CYFRA21-1) is one of the tumor markers used in diagnosing NSCLC. Type I and type II CKs have been shown to form obligate 1:1 heteropolymers, suggesting that dynamic changes must occur in the expression levels of CK pools when one CK is suppressed. However, the absolute levels of CK expression and their dynamic changes have not been fully evaluated. Therefore, we quantitatively determined CK expression levels in NSCLC cell lines, and evaluated the rate of change of CK expression levels after RNA interference targeting of single CKs. In NSCLC cells, type I CK18 and type II CK8 are the dominant CKs, with absolute expression levels of 12-77pmol/10(6)cells, while the expression patterns of the CKs vary among cell lines. Moderate suppression of a single dominant CK caused downregulation in CKs of the complementary type, and upregulation of other CKs of the same type. In contrast, severe suppression of a single dominant CK caused almost complete suppression of all CKs. In addition, introduction of CK19 led to resistance to CK degradation by CK18 suppression. These data suggest the presence of a critical threshold expression level for a dominant CK and a role for CK19 in the compensation of type I and type II CK pools in NSCLC.