Two-dimensional polyacrylamide gel electrophoresis analysis of indomethacin-treated human colon cancer cells.
OBJECTIVE
To establish the two-dimensional gel electrophoresis (2-DE) profiles of indomethacin (IN)-treated human colon cancer cell line HCT116, and to provide a new way to study its anti-tumor molecular mechanism through analyzing a variety of protein maps.


METHODS
Two-DE profiles of HCT116 were established in IN-treated and untreated groups. Total proteins were separated by immobilized pH gradient-based 2-DE. The gels were stained by silver, scanned by ImageScanner, and analyzed with Image Master software.


RESULTS
Clear background, well-resolved and reproducible 2-DE patterns of HCT116 cells were acquired in IN-treated and untreated group. The average deviation of spot position was 0.896+/-0.177 mm in IEF direction and 1.106+/-0.289 mm in SDS-PAGE direction respectively. In IN-treated group, 1169+/-36 spots were detected and 1061+/-32 spots were matched, the average matching rate was 90.6% in three gels. In untreated group, 1256+/-50 spots were detected and 1168+/-46 spots were matched, the average matching rate was 93.0% in three gels. Forty-five differential protein spots were displayed between IN-treated and untreated groups. Of which, 34 protein spots decreased and 9 showed higher expression in IN-treated group, and only two protein spots showed an expression in untreated cells.


CONCLUSIONS
Two-DE profiles of IN-treated and untreated HCT116 cells were established. Apparent 45 different protein spots were detected in IN-treated and untreated HCT116 cells. The analysis on differential protein spots may serve as a new way to study the molecule mechanism of IN-treated colon cancer.
Cheng
Yan-Li
YL
Department of Gastroenterology, Xiangya Hospital, Central South University, Xiangya Road, Changsha 410008, Hunan Province, China.
Zhang
Gui-Ying
GY
Xiao
Zhi-Qiang
ZQ
Tang
Fa-Qing
FQ
D000970:Antineoplastic Agents / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000737:chemistry* / Q000188:drug therapy*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods* / Q000592:standards; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology*; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results
D000970:Antineoplastic Agents; D007213:Indomethacin