Diagnosis of oesophageal cancer by detection of minichromosome maintenance 5 protein in gastric aspirates

Abstract

Symptomatic oesophageal cancer is usually advanced and the prognosis poor. Lethality of symptomatic oesophageal cancer has motivated screening for these diseases earlier in their evolution, but reliable methods for early diagnosis remain elusive. We have demonstrated that dysregulated expression of minichromosome maintenance (MCM) proteins 2 -7 is characteristic of early epithelial carcinogenesis, and that these key DNA replication initiation factors can be used as diagnostic markers for cervical and genito-urinary tract cancer. In this study, we investigated whether minichromosome maintenance protein 5 (Mcm5) can be used to detect oesophageal cancer cells in gastric aspirates. Two monoclonal antibodies raised against His-tagged human Mcm5 were used in a timeresolved immunofluorometric assay to measure Mcm5 levels in cells isolated from gastric aspirates of 40 patients undergoing gastroscopy for suspected or known oesophageal carcinoma or symptoms of dyspepsia. The test discriminated with high specificity and sensitivity between patients with and without oesophageal cancer (85% sensitivity (95% confidence interval (CI) ¼ 62 -97%), 85% specificity (CI ¼ 66 -96%)), as demonstrated by the large area under the receiver operating characteristics curve (0.93 (95% CI ¼ 0.85 -0.99)). Elevated levels of Mcm5 in gastric aspirates are highly predictive of oesophageal cancer. This simple test for oesophageal cancer is readily automated with potential applications in primary diagnosis, surveillance and screening.

The cell cycle and cancer

“…Notably, the arrested differentiation that characterizes cancer, particularly in high-grade tumours, is associated with failure to down-regulate the replication initiation proteins. (B) Spatial organization of Mcm2-7 protein expression in normal oesophageal squamous epithelium and non-dysplastic Barrett's mucosa and disruption of this highly organized spatial arrangement in premalignant dysplasia and invasive cancer [69,96]. In normal squamous epithelium, high expression levels of Mcm2 above a well-defined basal layer fall to undetectable levels in the upper third.…”

“…The detection of exfoliated MCM positive cells in body fluids (eg in urine, prostatic secretions, stool samples or gastro-oesophageal aspirates) or active sampling by swabbing or brushing (eg cervical smears or ERCP brushings for pancreaticobiliary tract sampling) provides a sensitive and specific method for the detection of premalignant and malignant lesions in a range of organ systems [39,[91][92][93][94][95][96][97][98]. For example, the immunostaining of cervical Pap smears for Mcm2-7 has potential to increase both the sensitivity and specificity of this error-prone test [39,[99][100][101].…”

“…Using this approach to study breast cancer, a complex and highly heterogeneous tumour type with respect to cancer genetics and clinicopathological parameters, has revealed three discrete cell cycle phenotypes [110,111] (Figure 3B). These include: (a) an out-ofcycle state composed predominantly of MCM-negative cells; (b) a G 1 -delayed/arrested state composed of cells The receiver operating characteristics curves show the high sensitivity and specificity of MCM-based tests for detection of oesophageal, bladder and prostate cancer [95,96,98], while the detection of (pre)malignant cells in cervical smears is illustrative of the potential for cytology-based testing [39,93].…”

Cell‐cycle‐dependent regulation of DNA replication and its relevance to cancer pathology

“…In this case, the expansion of the proliferative compartment with increasing histological grade in preinvasive lesions is reflected by the appearance of MCM-positive abnormal cells at the surface of the epithelium. The dysregulated pattern of MCM expression has been detected in dysplasia and malignancy of cervical [14,15], laryngeal [16], colorectal [17,18], oesophageal [19][20][21], urothelial [22], and vulval [23] epithelia.…”

Diagnosis of pancreaticobiliary malignancy by detection of minichromosome maintenance protein 5 in bile aspirates

“…We have also shown that dysregulation of MCM proteins is an early event in epithelial carcinogenesis, which occurs in a wide range of preneoplastic and neoplastic states resulting in exfoliation of MCM-positive tumour cells. Moreover, we have utilised these novel biomarkers of growth as diagnostic markers of cervical, genitourinary tract and oesophageal cancer (Williams et al, , 2004Stoeber et al, 1999Stoeber et al, , 2002.…”