Korean J Gynecol Oncol Colposc.  2004 Jun;15(2):113-122.

Proteomic Analysis of Anti-Proliferative Effects by Treatment of 5-Fluorouracil in Cervical Cancer Cells

• Kim CJ
• Yim EK
• Lee KH
• Ro DY
• Bae JS
• Park TC
• Namkoong S
• Um SJ
• Park JS

Abstract

OBJECTIVE
We investigated the effects of 5-fluorouracil (FU) to the cervical cancer cell lines with HPV in the aspect of the apoptosis and HPV.
METHODS
The effects of 5-FU on apoptosis was studied using a proteomic method based on a two dimensional gel electrophoresis and MALDI-TOF-MS using peptide mass fingerprinting.
RESULTS
More than 50 proteins showed a significant change in 5-FU treated cervical carcinoma cells compared to control cells. Among them, 34 proteins have been identified and 22 proteins were up-regulated CIDE-B (cell death inducing DFFA-like effector B), caspase-3, caspase-8 and Apo-1/CD95 (Fas), etc and 12 proteins were down-regulated mitotic checkpoint protein BUB3, myc proto-oncogene protein (c-myc), src substrate cortactin and transforming protein p21A, etc by 5-FU treatment in HeLa cells as determined by spot volume (P<0.05). Our experiments showed that 5-FU engaged the mitochondrial apoptotic pathway involving cytosolic cytochrome c release and subsequent activation of caspase-9 and caspase-3 as well as the membrane death receptor (DR)-mediated apoptotic pathway involving activation of caspase-8 with an Apo-1/CD95 (Fas)-dependent fashion. In addition, we could observe reduction of HPV-18 E6/E7 gene expression and activation of p53, pRb and p21(waf1) proteins by 5-FU treatment in HeLa cells.
CONCLUSION
we suggest that 5-FU suppresses the growth of cervical cancer cells not only by anti-proliferative effect but also anti-viral regulation. Our findings may offer new insights into the mechanism of anticancer effect affected by 5-FU treatment in cervical cancer cells and its mode of action.

Keyword

5-Fluorouracil; Apoptosis; HPV-18 E6/E7; MALDI-TOF-MS

MeSH Terms

Apoptosis
Caspase 3
Caspase 8
Caspase 9
Cell Line
Cortactin
Cytochromes c
Cytosol
Dermatoglyphics
Electrophoresis, Gel, Two-Dimensional
Fluorouracil*
Gene Expression
HeLa Cells
Human papillomavirus 18
Humans
M Phase Cell Cycle Checkpoints
Membranes
Proto-Oncogenes
Uterine Cervical Neoplasms*
Caspase 3
Caspase 8
Caspase 9
Cortactin
Cytochromes c
Fluorouracil