Isodon eriocalyx and its bioactive component Eriocalyxin B enhance cytotoxic and apoptotic effects of gemcitabine in pancreatic cancer.

BACKGROUND: Pancreatic cancer, associated with poor prognosis and low survival rate, has been the fourth leading cause of cancer-related death in the US. Although gemcitabine (Gem) is the first-line chemotherapeutic drug in the management of pancreatic cancer, the median survival extension is only 1.5 months, indicating unsatisfactory clinical results. Therefore, exploring agents that can enhance the anti-cancer activity of Gem would be an attractive strategy. PURPOSE: Our previous studies have demonstrated that eriocalyxin b (EriB), an ent­kaurane diterpenoid isolated from Isodon eriocalyx (Dunn.) Hara, possesses anti-pancreatic cancer effects, thus acting as a potential therapeutic agent. In this study, we further investigated whether EriB or the ethanol extract of I. eriocalyx (Isodon) could potentiate the cytotoxic activity of Gem in human pancreatic adenocarcinoma cells. In addition, the mechanism associated with their effects was also studied. METHODS: The anti-proliferation effect was assessed by MTT assay and Ki-67 immunostaining. The combination effect (addition, synergism and antagonism) of various agents was calculated by the Calcusyn software (Biosoft), utilizing the T.C. Chou Method. Apoptosis was detected using Annexin V and PI double staining followed by quantitative flow cytometry. Protein expression regulated by various treatments was analyzed by western blotting. RESULTS: The combination index revealed that Gem and EriB (or Isodon extract) had synergistic anti-proliferative effect. Both cellular apoptotic and anti-proliferative effects of Gem were significantly increased after combination with EriB (or Isodon extract). The underlying mechanisms involved in the combination effects were elucidated, which include: (1) increased activation of the caspase cascade; (2) reduction of PDK1 and AKT phosphorylation; (3) induction of JNK phosphorylation by Isodon and Gem combination. CONCLUSION: Gem and EriB (or Isodon extract) taken together in combination regulated PDK1/AKT1/caspase and JNK signaling and promoted apoptosis synergistically, which may contribute to the much increased anti-proliferative activity compared to either agent alone.

Eriocalyxin B-induced apoptosis in pancreatic adenocarcinoma cells through thiol-containing antioxidant systems and downstream signalling pathways.

Thiol-containing antioxidant systems play an important role in regulating cellular redox homeostasis. Several anti-cancer agents act by targeting these systems by inducing the production of reactive oxygen species (ROS). Our earlier studies have shown that Eriocalyxin B (EriB), a diterpenoid isolated from Isodon eriocalyx, possesses anti-pancreatic tumour activities in vitro and in vivo. The present study further demonstrated that only thiol-containing antioxidants, N-acetylcysteine (NAC) or dithiothreitol (DTT), inhibited EriB-induced cytotoxicity and apoptosis. EriB suppressed the glutathione and thioredoxin antioxidant systems, thus increasing the intracellular ROS levels and regulating the MAPK, NFκB pathways. Treatment with EriB depleted the intracellular thiol-containing proteins in CAPAN-2 cells. In vivo studies also showed that EriB treatment (2.5 mg/kg) reduced the pancreatic tumour weights significantly in nude mice with increased superoxide levels. Taken together, our results shed important new light on the molecular mechanisms of EriB acting as an apoptogenic agent and its therapeutic potential for pancreatic cancer.

Longikaurin A, a natural ent-kaurane, induces G2/M phase arrest via downregulation of Skp2 and apoptosis induction through ROS/JNK/c-Jun pathway in hepatocellular carcinoma cells.

Hepatocellular carcinoma (HCC) is the most common form of primary liver cancer, and is also highly resistant to conventional chemotherapy treatments. In this study, we report that Longikaurin A (LK-A), an ent-kaurane diterpenoid isolated from the plant Isodon ternifolius, induced cell cycle arrest and apoptosis in human HCC cell lines. LK-A also suppressed tumor growth in SMMC-7721 xenograft models, without inducing any notable major organ-related toxicity. LK-A treatment led to reduced expression of the proto-oncogene S phase kinase-associated protein 2 (Skp2) in SMMC-7721 cells. Lower Skp2 levels correlated with increased expression of p21 and p-cdc2 (Try15), and a corresponding decrease in protein levels of Cyclin B1 and cdc2. Overexpression of Skp2 significantly inhibited LK-A-induced cell cycle arrest in SMMC-7721 cells, suggesting that LK-A may target Skp2 to arrest cells at the G2/M phase. LK-A also induced reactive oxygen species (ROS) production and apoptosis in SMMC-7721 cells. LK-A induced phosphorylation of c-Jun N-terminal kinase (JNK), but not extracellular signal-regulated kinase and P38 MAP kinase. Treatment with, the JNK inhibitor SP600125 prevented LK-A-induced apoptosis in SMMC-7721 cells. Moreover, the antioxidant N-acetylcysteine prevented phosphorylation of both JNK and c-Jun. Taken together, these data indicate that LK-A induces cell cycle arrest and apoptosis in cancer cells by dampening Skp2 expression, and thereby activating the ROS/JNK/c-Jun signaling pathways. LK-A is therefore a potential lead compound for development of antitumor drugs targeting HCC.

A new technology for harnessing the dye polluted water and dye collection in a chemical factory.

A new technology for harnessing the dye polluted water and dye collection was developed. It is based on the enhanced evaporation by using solar, wind and air temperature energy and additional heat-electric energy. It consists of four parts: (1) evaporation carrier system (evaporation carrier and frame for evaporation carrier) for polluted water; (2) polluted water circulating system (pumping-spraying-collecting); (3) heating system; (4) workshop with polluted water reservoir-tanks and rainfall prevention roof. The polluted water was (heated in case necessary) sprayed to the evaporation carrier system and the water was evaporated when it moved in the space and downward along the carrier mainly by using natural (solar, wind and air temperature energy). In case, when there is no roof for the carrier system, the polluted water can be stored in the reservoirs (storage volume for about 20 days). The first 10-25 mm rainfall also need to be stored in the reservoirs to meet the state standard for discharging wastewater. The dye may be collected at the surface in the reservoir-tanks and the crystallized salt may be collected at the bottom plate. The black-color wastewater released by the factory is no more discharged to the surface water system of Taihu Lake Basin. About 2 kg dye and 200 kg industrial salt may be collected from each tone of the polluted water. The non-pollution production of dye may be realized by using this technology with environmental, economical and social benefits.

Suppressive effect of high energy shock waves on tumor cells.

Suppressive effect on tumor cells of high energy shock waves (HESW) has aroused the interest of physicians in recent years. We assessed experimentally the cytotoxic effects of HESW on tumor cells both in vitro and in vivo and determined whether a Chinese domestic lithotriptor is capable of generating effective HESW, which has the potential to break tumor cells, reduce cell viability, retard cell growth, delay doubling time, impair cell attachment and cell clonogenicity. In nude rats, HESW was able to delay tumor growth and reduce tumor size without evidence of metastasis. The nature of HESW in the induction of cell damage and its clinical application need to be further investigated.

[Study on dissolution rate of injection of microencapsulated compound megestrol acetate].

Dissolution rate of megestrol acetate in the injection of microencapsulated compound megestrol acetate was determined by the first derivative spectrum amplitude method. The experimental results reveal that t50 of the microencapsulated sample (I) is about 60.16 d, while the broken microencapsulated sample (II) is about 15.89 d and the unmicroencapsulated sample (III) about 15.87 d. The difference is regarded as of obvious significance (P less than 0.01). The values of the dissolution rate were linear with the parameters of rabbits and women both in vivo.