Detection of Cu(I) and Zn(II) ions in colon tissues by multi-photon microscopy: novel marker of antioxidant status of colon neoplasm.
OBJECTIVE
Establishing probe-based analysis is important for developing multi-photon microscopy (MPM) to make an early diagnosis of colon neoplasm and assess its antioxidant status. Cu(I) and Zn(II) ions are trace elements which roll as cofactors of antioxidant, superoxide dismutase. However, there have been no reports on the features of MPM image using probe of Cu(I) and Zn(II) ions. Our main objective in this study was application of newly developed multi-photon probe (MP) probe on Cu(I) and Zn(II) ions as a tool to assess antioxidative status of colon neoplasm.


METHODS
This study was a pilot study. Colon cancer cell lines (ACT116 and HT-29), NIH3T3 cells and tissues of normal colon mucosa and colon neoplasm obtained during colonoscopic biopsy from 17 patients were stained with MP probes for Cu(I) and Zn(II) (ACu1 and AZn1). Cu(I)/Zn(II) levels in the cells and tissues were determined by detecting MP-excited fluorescence by MPM.


RESULTS
MPM images of cells stained with MP probes revealed that Cu(I) was more abundant in ACT116 and HT-29 cells than in NIH3T3 cells, while Zn(II) was more abundant in NIH3T3 cells than in ACT116 and HT-29 cells. Normal tissues had a defined texture, whereas adenoma/adenocarcinoma tissues were amorphous. The level of Cu(I) increased and that of Zn(II) decreased with the transition from normal to adenoma to adenocarcinoma tissue.


CONCLUSIONS
MPM can be used to determine the relative Cu(I)/Zn(II) levels in cells and colon tissues by using ACu1 and AZn1 as MPM probes. These finding are new research tools for gastroenterologists to assess antioxidant status of colon neoplasm.
Division of Gastroenterology and Hepatology, Department of Internal Medicine, Institute of Nutritional and Digestive Disease, Korea University College of Medicine, Seoul, Korea.
Eun Sun
ES
Kim
Chang Su
CS
Lim
Hoon Jai
HJ
Chun
Bora
B
Keum
Yeon Seok
YS
Seo
Yong Sik
YS
Kim
Yoon Tae
YT
Jeen
Hong Sik
HS
Lee
Soon Ho
SH
Um
Chang Duck
CD
Kim
Ho Sang
HS
Ryu
Bong Rae
BR
Cho
D000328:Adult; D000368:Aged; D000975:Antioxidants / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism*; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000175:diagnosis*; D003300:Copper / Q000378:metabolism*; D006801:Humans; D007477:Ions / Q000378:metabolism; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D008875:Middle Aged; D015335:Molecular Probes / Q000378:metabolism; D010865:Pilot Projects; D014131:Trace Elements / Q000378:metabolism; D015032:Zinc / Q000378:metabolism*
D000975:Antioxidants; D014408:Biomarkers, Tumor; D007477:Ions; D015335:Molecular Probes; D014131:Trace Elements; D003300:Copper; D015032:Zinc