Camelliin B induced apoptosis in HeLa cell line

Gordonia axillaris (Roxb.) Dietrich (Theaceae) is a native to Taiwan and the leaves have been used as an astringent folk medicine. Camelliin B (CB), a macrocyclic hydrolyzable tannin, was isolated from G. axillaris and showed cytotoxic effects in human carcinoma cells. Among the target cells (SKHep-1, Ha-22T, DU-145, AGS, and HeLa), the cervical carcinoma cell line, HeLa, was more sensitive to CB than were Chang normal liver cells and primary-cultured normal gingival and cervical fibroblasts. Furthermore, the cytotoxic effects of CB showed dose-dependency at 3.2-100.0 microg/ml in HeLa for 1,24,48, and 72 h and with an IC(50) value of 46.3 microg/ml for 48 h. However, the IC(50) value of CB in primary-cultured normal cervical fibroblasts was 108.0 microg/ml. Therefore, the selectivity shown by CB was ascribed to the difference in growth speed between normal and tumor cells. HeLa cells and primary-cultured normal cervical fibroblasts were treated with 50.0 and 100.0 microg/ml CB for 48 h, respectively, and exhibited chromatin condensation, indicating the occurrence of apoptosis. Flow cytometric analysis demonstrated the presence of apoptotic cells with low DNA content, a decrease of cell population at the G(1) phase, and a concomitant increase of cell population at the G(2)/M phase. CB also caused DNA fragmentation and inhibited PARP degradation in HeLa cells. However, CB did not significantly inhibit Bcl-2 expression in HeLa cells at 50.0 microg/ml, only at 100.0 microg/ml for 48 h. These results suggest that CB induced apoptosis, without direct inhibition of Bcl-2 expression in HeLa cells.