title: Sequential treatment with betulinic acid followed by 5-fluorouracil shows synergistic cytotoxic activity in ovarian cancer cells.
abstract: Betulinic acid selectively inhibits the growth of ovarian carcinoma cell lines without affecting the normal cells. In the present study, the effect of 5-fluorouracil (5-FU) and betulinic acid (BA) combination on ovarian carcinoma cells was studied. The results demonstrated that ovarian carcinoma cells on concurrent or 5-FU followed by BA treatment show increased Sub-G1 cell population, increased rate of cell apoptosis and morphological changes in mitochondrial membrane. In OVCAR 432 cells treatment with sequential combination of 5-FU and BA increased the Sub-G1 cell population to 51.3% and growth inhibition rate of > 72%. However, exposure to BA before 5-FU treatment caused a decrease in rate of inhibition to < 35%. Treatment with combination of 5 μM of 5-FU and 1 μM of BA for 48 h, led to an induction of apoptosis in 79.7% and induced morphological changes in OVCAR 432 cells. The Western blot results showed high concentration of cytochrome c in the cell cytosol after 24 h of 5-FU and BA combination treatment. Treatment of BA-responsive RMS-13 cells with 5-FU and BA combination resulted in inhibition of GLI1, GLI2, PTCH1, and IGF2 genes. In addition, we found a significant reduction in hedgehog activity of RMS-13 cells after 5-FU and BA combination treatment by means of a hedgehog-responsive reporter assay. Therefore, 5-FU and BA combination can be a promising regimen for the treatment of ovarian carcinoma.
authors.affiliation: Department of Obstetrics and Gynecology, China-Japan Union Hospital of Jilin University Changchun 130000, China.
authors.forename: Ying-Jian
authors.initials: YJ
authors.lastname: Wang
authors.affiliation: Department of Obstetrics and Gynecology, China-Japan Union Hospital of Jilin University Changchun 130000, China.
authors.forename: Jun-Bao
authors.initials: JB
authors.lastname: Liu
authors.affiliation: Department of Traditional Chinese Medicine, China-Japan Union Hospital of Jilin University Changchun 130000, China.
authors.forename: Yu-Chang
authors.initials: YC
authors.lastname: Dou
mesh_terms: D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002277:Carcinoma / Q000473:pathology*; D045744:Cell Line, Tumor; D004357:Drug Synergism; D005260:Female; D005434:Flow Cytometry; D005472:Fluorouracil / Q000008:administration & dosage; D053823:Hedgehog Proteins / Q000378:metabolism; D006801:Humans; D010051:Ovarian Neoplasms / Q000473:pathology*; D053978:Pentacyclic Triterpenes; D014315:Triterpenes / Q000008:administration & dosage; D000094062:Betulinic Acid
chemical_list: D053823:Hedgehog Proteins; D053978:Pentacyclic Triterpenes; D014315:Triterpenes; D005472:Fluorouracil; D000094062:Betulinic Acid
keywords: Ovarian carcinoma; apoptosis; hedgehog; inhibition; morphological changes