Improved screening for anal neoplasia by immunocytochemical detection of minichromosome maintenance proteins.
OBJECTIVE
Early detection of anal intraepithelial neoplasia (AIN) and anal squamous cell carcinoma (SCC) by screening will improve clinical outcome. Assessment of anal cytology samples using routine Papanicolaou testing suffers from shortcomings in sensitivity and/or specificity, suggesting that screening tests based on biomarkers may be of value. We tested the suitability in this context of minichromosome maintenance (MCM) proteins, accurate markers of the deregulated cell cycle entry that characterizes malignancy and premalignancy.


METHODS
We undertook an initial immunohistochemical study of 54 anal tissue samples and validated our findings using an independent prospective cohort study of 235 anal cytology samples from 144 subjects.


RESULTS
In the progression from normal anal epithelium through AIN to SCC, there was increasing expression of MCM2 and MCM5, including in the superficial epithelial third, the source of the majority of cells collected by anal swab. The median labeling indices (LI) for MCM2 and MCM5 in the superficial third of AIN2/3 and SCCs combined were 90.2% and 84.0%, respectively. MCM LIs in the superficial layers were significantly greater than LIs for Ki67, an alternative marker of cell cycle entry (P<0.0001). By immunocytochemistry using a mixture of anti-MCM2 and anti-MCM5 antibodies, immunopositive cells were readily identified in anal cytology samples, even at low magnification. MCM testing showed sensitivity for AIN2/3 of 84% (95% confidence interval, 75,93) and for AIN1/viral changes of 76% (68, 84), with overall specificity (for any lesion) of 77% (64, 90).


CONCLUSIONS
MCMs are promising biomarkers for improving detection of AIN and SCC in anal cytology samples.
Medical Research Council Cancer Cell Unit, Addenbrooke's Hospital, Cambridge, United Kingdom.
Cinzia
C
Scarpini
Victoria
V
White
Balaji
B
Muralidhar
Aileen
A
Patterson
Nicola
N
Hickey
Naveena
N
Singh
Jose
J
Mullerat
Marc
M
Winslet
R Justin
RJ
Davies
Marie-Louise
ML
Phillips
Pamela
P
Stacey
Ronald A
RA
Laskey
Richard
R
Miller
Mayura
M
Nathan
Nicholas
N
Coleman
D000328:Adult; D000368:Aged; D001005:Anus Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism*; D018450:Disease Progression; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D064111:Minichromosome Maintenance Complex Component 2; D009687:Nuclear Proteins / Q000378:metabolism*; D011446:Prospective Studies; D012680:Sensitivity and Specificity
D018797:Cell Cycle Proteins; D019394:Ki-67 Antigen; C489423:MCM5 protein, human; D009687:Nuclear Proteins; D064111:Minichromosome Maintenance Complex Component 2