RESUMEN
Hepatic cancer remains a challenging disease and there is a need to identify new treatments. Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional medicinal prescription, has been used to treat lymphadenopathy and exhibits cytotoxic activity in many types of human cancer cells. Our previous studies revealed that SJKJT is capable of inhibiting colon cancer colo 205 cells by inducing autophagy and apoptosis. However, the effects and molecular mechanisms of SJKJT in human hepatocellular carcinoma have not been clearly elucidated. In the present study we evaluated the effects of SJKJT in human hepatic cellular carcinoma Hep-G2 cells. The cytotoxicity of SJKJT in Hep-G2 cells was measured by MTT assay. The cell cycles were analyzed by fluorescenceactivated cell sorting (FACS). The protein expression of translationally controlled tumor protein (TCTP), Mcl-1, Fas, TNF-α, Caspase-8, Caspase-3 and Bax in Hep-G2 cells treated with SJKJT was evaluated by western blotting. The protein expression of Caspase-3 was also detected by immunofluorescence staining. The results showed that SJKJT inhibits Hep-G2 cells in a time- and dosedependent manner. During SJKJT treatment for 48 and 72 h, the half-maximum inhibitory concentration (IC50) was 1.48 and 0.94 mg/ml, respectively. The FACS results revealed that increased doses of SJKJT were capable of increasing the percentage of cells in the sub-G1 phase. Immunofluorescence staining showed that Hep-G2 treated with SJKJT had increased expression of Caspase-3. The western blot results showed that the protein expression of Fas, TNF-α, Caspase-8, Caspase- 3 and Bax was upregulated, but that of TCTP and Mcl-1 was downregulated in Hep-G2 cells treated with SJKJT. In conclusion, these findings indicated that SJKJT inhibits Hep-G2 cells. One of the molecular mechanisms responsible for this may be the increased Fas, TNF-α, Caspase-8, Caspase- 3 and Bax expression; another mechanism may be via decreasing TCTP and Mcl-1 expression in order to induce apoptosis.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Carcinoma Hepatocelular/patología , Caspasas/metabolismo , Medicamentos Herbarios Chinos/uso terapéutico , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Apoptosis/efectos de los fármacos , Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/enzimología , Caspasa 3/metabolismo , Caspasa 8/metabolismo , Ciclo Celular/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Medicamentos Herbarios Chinos/farmacología , Proteína de Dominio de Muerte Asociada a Fas/metabolismo , Células Hep G2 , Humanos , Inmunohistoquímica , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/enzimología , Neoplasias Hepáticas/patología , Proteína 1 de la Secuencia de Leucemia de Células Mieloides , Proteína Tumoral Controlada Traslacionalmente 1 , Receptor fas/metabolismoRESUMEN
Tanshinone IIA (Tan-IIA) is extracted from Danshen, Salviae miltiorrhizae Radix, which has been widely adopted in traditional herbal medicine to treat cardiovascular and hepatic diseases. Tan-IIA induces apoptosis and inhibits proliferation in human hepatocellular carcinoma (HCC) cells. However, the molecular mechanisms of Tan-IIA on human HCC cells are not understood clearly. In the present study, the cytotoxicity of Tan-IIA as well as its molecular mechanisms in human HCC J5 cells was investigated. The cytotoxicity was assayed by MTT. The protein expression of p53, p21, Bax, Bcl-2, Cdc25c, Cdc2, calreticulin, caspase 12, GADD153, caspase 3 and beta-actin in J5 cells were determined by Western blotting. The cell cycles were analyzed by FACS. The protein expression of caspase 12, GADD1533 and caspase 3 were detected by immunocytochemical staining. The results showed that Tan-IIA inhibited J5 cells in a dose- and time-dependent manner. The protein expression of p53, p21, Bax, calreticulin, caspase 12, caspase 3 and GADD153 were increased, but Bcl-2, Cdc25c and Cdc2 were decreased in J5 cells. In addition, the results also showed that Tan-IIA arrested J5 cells in the G2/M phase. Immunocytochemistry staining showed that J5 cells treated with Tan-IIA up-regulated the protein expression of caspase 12, 3 and GADD153. Taken together, the findings suggest that Tan-IIA inhibits and induces apoptosis in J5 cells through novel molecular targets, calreticulin, caspase 12 and GADD153.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Calreticulina/metabolismo , Carcinoma Hepatocelular/tratamiento farmacológico , Caspasa 12/metabolismo , Neoplasias Hepáticas/tratamiento farmacológico , Fenantrenos/farmacología , Factor de Transcripción CHOP/metabolismo , Abietanos , Clorometilcetonas de Aminoácidos/metabolismo , Animales , Antineoplásicos Fitogénicos/uso terapéutico , Apoptosis/efectos de los fármacos , Inhibidores de Caspasas , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Inhibidores de Cisteína Proteinasa/metabolismo , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Humanos , Masculino , Mitocondrias/metabolismo , Fenantrenos/uso terapéuticoRESUMEN
Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional Chinese medicine prescription, has been used to treat lymph node diseases and tumors. However, the molecular mechanisms of SJKJT in human colon cancer in vivo and in vitro have not been clearly elucidated. In the present study, we investigated the molecular mechanisms of SJKJT in human colon cancer colo 205 cells in vitro and in vivo. In the in vitro study, colo 205 cells were treated with various concentrations (0.5, 1 and 2 mg/ml) of SJKJT. The protein expression of TNF-α, Caspase-8 and Caspase-3 in colo 205 cells was measured by Western blotting. The results demonstrate that SJKJT up-regulated Fas, TNF-α, Caspase-8 and Caspase-3 protein expression. In the in vivo study, human colon cancer colo 205 cells (3x106/0.2 ml) were injected subcutaneously into the flank area of nude SCID mice (n=32) randomly divided into four groups. SJKJT was dissolved in saline and then administered orally to the mice at concentrations of 0.01, 0.1 and 0.3 g/kg/day for 30 days. The control group was treated with an equal volume of saline. SCID mice were sacrified by CO2 inhalation and the xenograft tumors were dissected. Subsequently, the protein expression of Fas, TNF-α, Caspase-8 and Caspase-3 in the tumors was measured by Western blotting. The results demonstrate that SJKJT up-regulated Fas, TNF-α, Caspase-8 and Caspase-3 protein expression, both in vitro and in vivo. These observations suggest that SJKJT has therapeutic potential in colon cancer.
RESUMEN
Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional Chinese medicine, has been prescribed as complementary medication for colon cancer in Taiwan. However, its molecular mechanisms are not yet understood. In the present study, we investigated the effects of SJKJT on human colon cancer colo 205 cells in vitro. The cytotoxicity of SJKJT in colo 205 cells was evaluated using the MTT assay, and the protein expression of microtubule-associated protein II light chain 3 (MAP-LC3-II) was measured using Western blot analysis. The results showed that SJKJT inhibited the survival rates of colo 205 cells in a time- and dose-dependent manner, with an IC50 concentration at 24 h of 590.34 µg/ml. In addition, SJKJT up-regulated the protein expression of MAP-LC3-II in colo 205 cells. These findings indicate that one of the molecular mechanisms by which SJKJT inhibits the proliferation of colo 205 cells in?vitro may be through the induction of the autophagic pathway. SJKJT may therefore have therapeutic potential for the treatment of human colon cancer.
RESUMEN
The aim of this study was to evaluate peripheral nerve regeneration across a 15-mm gap in the sciatic nerve of the rat, using a silicone rubber nerve guide filled with different concentrations of astragaloside (0, 50, 100, and 200 microM). Collagen was also filled in the chambers to prevent the astragaloside from leakage. At the end of 8 weeks, animals from the group treated with astragaloside, especially at the concentration of 50 microM, had a higher rate of successful regeneration across the wide gap, a significantly larger number of myelinated axons, and a greater evoked action potential than the control group. However, the high-dose astragaloside (200 microM) completely reversed this positive effect of growth-promoting capability and inhibited nerve regeneration. Thus, astragaloside plays a dual role in anastomosis, being salutary in aiding the growth of axons in peripheral nerve but also detrimental, terminating the nerve regenerative processes if improperly applied.
Asunto(s)
Axones/fisiología , Regeneración/efectos de los fármacos , Saponinas/farmacología , Nervio Ciático/fisiología , Triterpenos/farmacología , Animales , Relación Dosis-Respuesta a Droga , Ratas , Ratas Sprague-Dawley , Regeneración/fisiología , Nervio Ciático/citologíaRESUMEN
The present study provides in vivo trials of silicone rubber chambers filled with different concentrations of bilobalide (0, 50, 100, 200, 400 microM) to bridge a 15 mm sciatic nerve defect in rats. Collagen was also filled in the chambers to prevent the bilobalide from leakage. Histological and electrophysiological techniques were used to evaluate the functional recovery of the nerve. At the conclusion of 8 weeks, animals from the group treated with the bilobalide, especially at the concentration of 200 microM, had a higher rate (40%) of successful regeneration across the wide gap and a significantly larger number of myelinated axons (4094 +/- 1555), compared to only 10% and 2485 in the control group. However, the high dose bilobalide (400 microM) completely reversed this positive effect of growth-promoting capability and inhibited nerve regeneration. Only 10% of the animals treated with the high dose bilobalide had regenerated cables within the silicone rubber chambers. These results indicated that bilobalide could be involved in both positive and negative effects on regenerating nerves. Therefore, whether a proper dosage of bilobalide is used or not plays a critical factor in deciding if it can sustain nerve regeneration over long gaps.