Korean J Obstet Gynecol.  2007 Mar;50(3):476-485.

Expression analysis of mRNA and proteins related to angiogenesis following management with cytotoxic agents in gynecologic cancer cell lines

• Kim YJ
• Kong MS
• Lee YM
• Chang SY
• Kim SR
• Kim KT
• Hwang YY
• Cho SH

Abstract

OBJECTIVE
Angiogenesis is central role to both the proliferation and metastasis of malignant tumor. The intense interest in angiogenesis has also lead to a re-examination of the activity of established cytotoxic agents which are known to be an antiangiogenic effect anecdotally. In this study, anti-angiogenic effect of cisplatin, paclitaxel and thalidomide was evaluated in human ovarian cancer cell lines and cervical cancer cell line.
METHODS
Human ovarian cancer cell line A2780, cisplatin resistant human ovarian cancer cell line A2780-CDDP, human breast cancer cell line MCF-7, and squamous cell uterine cervical carcinoma cell line SiHa were used to evaluate the level of mRNA and protein expression of VEGF, bFGF and TSP-1, 2 before and after the treatment with cisplatin, paclitaxel, and thalidomide using RT-PCR, protein extraction, and Western blot. The results were analyzed with Wilcoxon signed rank test in the SAS ver 8.1.
RESULTS
Targeted mRNAs were synthesized as 212 bp VEGF, 238 bp bFGF, and 492 bp band sized except mRNA of TSP-2 via RT-PCR. The protein of VEGF and bFGF were appeared as 21KDa and 17 KDa size, however, the protein of TSP-1 was not appeared through western blot. No effect of cisplatin on protein expression was measured in these cell lines, but paclitaxel influenced the expression of bFGF in MCF-7 cell line and the expression of TSP-1 in MCF-7 and SiHa cell lines. TSP-1 expression was influenced by thalidomide in A2780 cell line. The protein expression of VEGF and bFGF were not influenced following treatment with cisplatin, paclitaxel, and thalidomide.
CONCLUSION
These results were suggested that bFGF and TSP-1 will be used as a target gene for the assay of antiangiogenic effect of paclitaxel in breast and uterine cervical cancer tissue and TSP-1 will be used as that of thalidomide in ovarian cancer. Furthermore, thalidomide will be tried as an adjunctive agent for the improvement of the survival in the case of the patient with ovarian cancer.

Keyword

Gynecologic cancer; Angiogenesis; Cisplatin; Paclitaxel; Thalidomide

MeSH Terms

Blotting, Western
Breast
Breast Neoplasms
Cell Line*
Cisplatin
Cytotoxins*
Humans
MCF-7 Cells
Neoplasm Metastasis
Ovarian Neoplasms
Paclitaxel
RNA, Messenger*
Thalidomide
Thrombospondin 1
Uterine Cervical Neoplasms
Vascular Endothelial Growth Factor A
Cisplatin
Cytotoxins
Paclitaxel
RNA, Messenger
Thalidomide
Thrombospondin 1
Vascular Endothelial Growth Factor A