Pancreatic cancer in vitro toxicity mediated by Chinese herbs SPES and PC-SPES: implications for monotherapy and combination treatment.

One of the greatest challenges in the treatment of pancreatic cancer remains its inherent lack of beneficial response to cytotoxic chemotherapy. Chinese herbal extracts have been widely used for the treatment of various cancers, but objective information on their efficacy in pancreatic cancer is lacking. Eight human pancreatic cancer cell lines (MIA, Panc-1, BxPC, ASPC, HS-766T, CaPan-2, CFPAC, and HTB-147) were studied for in vitro susceptibility to ethanol extracts of SPES and PC-SPES, two quality-controlled, dried, encapsulated supplements of 15 and eight Chinese herbs, respectively. Resulting toxicities, alone and in combination with doxorubicin or gemcitabine, were analyzed by [(3)H]thymidine incorporation or sulforhodamine B staining, colony formation, and TUNEL flow cytometry assays. Combination toxicity mechanisms were calculated by the combination index method of Chou and Talalay. In all cell lines, there was dose-dependent inhibition of proliferation. By [(3)H]thymidine incorporation assay, 50% growth inhibition after 48 h continuous exposure (IC(50)) occurred at concentrations of 0.2-0.8 microl/ml SPES and 0.4-1.3 microl/ml PC-SPES. Growth inhibition was accompanied by a significant enhancement of the TUNEL-positive apoptotic fraction of all cell lines after treatment with either extract. After treatment with PC-SPES, the cell lines consistently displayed a G2 cell cycle block; SPES induced an increase in S phase, with a smaller impact on G2. When added at a concentration of 0.2 microl/ml (approximately IC(20)), both extracts enhanced Panc-1 cell killing mediated by doxorubicin, with an average decrease in the corresponding IC(50) of 33% (range 11-62%). Combination effects with either extract appeared to be antagonistic in the case of gemcitabine and additive to mildly synergistic in the case of doxorubicin. Both SPES and PC-SPES exhibited significant toxicity in pancreatic cancer cells, mediated via induction of apoptosis. Both mixtures should be evaluated for their in vivo and clinical therapeutic utility as monotherapy agents against pancreatic cancer. SPES could possibly be combined with cell cycle-independent cytotoxic drugs. Due to a consistent G2 blocking pattern, PC-SPES may prove useful as a radiation sensitizer.

Chemosensitization by fibroblast growth factor-2 is not dependent upon proliferation, S-phase accumulation, or p53 status.

Fibroblast growth factor-2 (bFGF/FGF-2) is a pleiotropic growth factor that functions as a survival factor and directs apoptosis during embryogenesis and development. As a survival factor, FGF-2 would be expected to protect cells against drug toxicities. Such protection has been reported in some cells treated with some chemotherapeutic drugs. However, we recently demonstrated that FGF-2 can sensitize NIH 3T3 mouse fibroblasts to the cytotoxic and apoptotic effects of cisplatin. Sensitization requires prolonged incubation of cells with FGF-2 before the addition of cisplatin, and it requires an FGF-2 concentration (5-10 ng/mL) that is higher than that needed for its mitogenic effects (0.5 ng/mL). We now report that FGF-2 can also sensitize MCF7 human breast cancer cells and A2780 human ovarian cancer cells, as well as NIH 3T3 cells, to cisplatin. FGF-2 did not affect the cisplatin sensitivity of SKOV3 ovarian cancer cells or a panel of seven pancreatic cancer cell lines. We have demonstrated that the sensitizing effect is not simply a function of the mitogenic activity of FGF-2 on cells, as we did not observe sensitization with other growth-stimulatory factors (FGF-1 and epidermal growth factor); the sensitizing effect of FGF-2 was observed even with cell lines that were not growth-stimulated by FGF-2; and sensitization was not restricted to cells in S-phase of the cell cycle. These results indicate that cell proliferation is neither necessary nor sufficient for sensitization by FGF-2. Moreover, sensitization to cisplatin appears to be p53-independent, as p53-null 3T3 10-1 cells were equally sensitized by FGF-2. Finally, FGF-2 also sensitized NIH 3T3 and MCF7 cells to carboplatin, and had smaller effects on the sensitivity of these cell lines to doxorubicin and docetaxel. FGF-2 had no effect on sensitivity to etoposide in any cell line tested. Therefore, sensitization by FGF-2 was most effective with the platinum compounds, suggesting that this activity may be specific to particular mechanisms of drug action.