Wang Ying-Jian, Liu Jun-Bao, Dou Yu-Chang
Department of Obstetrics and Gynecology, China-Japan Union Hospital of Jilin University Changchun 130000, China.
Department of Traditional Chinese Medicine, China-Japan Union Hospital of Jilin University Changchun 130000, China.
Int J Clin Exp Pathol. 2015 Jan 1;8(1):252-9. eCollection 2015.
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.
桦木酸可选择性抑制卵巢癌细胞系的生长，而不影响正常细胞。在本研究中，研究了5-氟尿嘧啶（5-FU）与桦木酸（BA）联合使用对卵巢癌细胞的影响。结果表明，同时使用5-FU和BA或先使用5-FU再使用BA处理的卵巢癌细胞，其亚G1期细胞群体增加，细胞凋亡率升高，线粒体膜出现形态变化。在OVCAR 432细胞中，5-FU和BA序贯联合处理使亚G1期细胞群体增加到51.3%，生长抑制率>72%。然而，在5-FU处理前先使用BA会导致抑制率降至<35%。用5μM的5-FU和1μM的BA联合处理48小时，导致OVCAR 432细胞中79.7%的细胞发生凋亡并出现形态变化。蛋白质印迹结果显示，5-FU和BA联合处理24小时后，细胞胞质溶胶中细胞色素c浓度升高。用5-FU和BA联合处理对BA敏感的RMS-13细胞，导致GLI1、GLI2、PTCH1和IGF2基因受到抑制。此外，通过刺猬信号通路响应报告基因检测，我们发现5-FU和BA联合处理后，RMS-13细胞的刺猬信号通路活性显著降低。因此，5-FU和BA联合使用可能是一种有前景的卵巢癌治疗方案。
