A new method for isolating colonocytes from naturally evacuated feces and its clinical application to colorectal cancer diagnosis.
OBJECTIVE
The early detection of colorectal cancer is desired because this cancer can be cured surgically if diagnosed early. The purpose of the present study was to determine the feasibility of a new methodology for isolating colonocytes from naturally evacuated feces, followed by cytology or molecular biology of the colonocytes to detect colorectal cancer originating from any part of the colorectum.


METHODS
Several simulation studies were conducted to establish the optimal methods for retrieving colonocytes from any portion of feces. Colonocytes exfoliated into feces, which had been retrieved from 116 patients with colorectal cancer and 83 healthy volunteers, were analyzed. Part of the exfoliated colonocytes was examined cytologically, whereas the remainder was subjected to DNA analysis. The extracted DNA was examined for mutations of the APC, K-ras, and p53 genes using direct sequence analysis and was also subjected to microsatellite instability (MSI) analysis.


RESULTS
In the DNA analysis, the overall sensitivity and specificity were 71% (82 of 116) of patients with colorectal cancer and 88% (73 of 83) of healthy volunteers. The sensitivity for Dukes A and B was 72% (44 of 61). Furthermore, the sensitivity for cancers on the right side of the colon was 57% (20 of 35). The detection rate for genetic alterations using our methodology was 86% (80 of 93) when the analysis was limited to cases in which genetic alterations were present in the cancer tissue.


CONCLUSIONS
We have developed a new methodology for isolating colonocytes from feces. The present study describes a promising procedure for future clinical evaluations and the early detection of colorectal cancers, including right-side colon cancer.
Matsushita
Hisayuki
H
Investigative Treatment Division, Research Center for Innovative Oncology, National Cancer Center Hospital East, Kashiwa, Japan.
Matsumura
Yasuhiro
Y
Moriya
Yoshihiro
Y
Akasu
Takayuki
T
Fujita
Shin
S
Yamamoto
Seiichiro
S
Onouchi
Shigeki
S
Saito
Norio
N
Sugito
Masanori
M
Ito
Masaaki
M
Kozu
Takahiro
T
Minowa
Takashi
T
Nomura
Sayuri
S
Tsunoda
Hiroyuki
H
Kakizoe
Tadao
T
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor; D003106:Colon / Q000166:cytology*; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology*; D004252:DNA Mutational Analysis; D005243:Feces / Q000166:cytology*; D005260:Female; D006801:Humans; D018189:Immunomagnetic Separation; D008297:Male; D008875:Middle Aged; D011446:Prospective Studies
D014408:Biomarkers, Tumor