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Fan J.,Zhejiang University | Wang J.-C.,The Xinhua Hospital | Chen Y.,Zhejiang University | Fang H.,The First Peoples Hospital of Taizhou | And 4 more authors.
African Journal of Biotechnology | Year: 2011

Sinomenine (SN) is an alkaloid that has previously been shown to have a role in such varied processes as inflammation, angiogenesis, arthritis and immunosuppression. Therefore, in this study, possible anti-tumor effects of SN on the human prostate cancer cell lines, PC-3 and DU-145, was investigated. After treatment with varying doses of SN, apoptosis was measured by flow cytometry. Prostaglandin E2 production was detected by ELISA, and expression levels of cyclooxygenase-2 and nuclear factorkappa B were evaluated by western blot. In both PC-3 and DU-145 cells, SN treatment induced apoptosis, inhibited in vitro production of prostaglandin E2, and decreased activation of cyclooxygenase-2 and nuclear factor-kappa B. Growth inhibition assays were also performed to determine whether cell viability was responsible for the decrease in the activation observed. In combination, these results suggest that SN can induce an anti-tumor response by suppressing the activation of cyclooxygenase-2 and nuclear factor-kappa B signaling pathways. © 2011 Academic Journals.

Huang Z.,Zhejiang University | Jin J.,Zhejiang University | Tong X.,Zhejiang University | Yang Q.,The Fuyang TCM Hospital | And 4 more authors.
Journal of Medicinal Plant Research | Year: 2011

Cordyceps sinensis (CS), a tonic herb in Chinese traditional medicine, is an excellent antitumor agent. However, the underlying mechanisms of its antitumor effects remain unclear. In the present study, we evaluated the role of C. sinensis in the differentiation and function of bone marrow-derived dendritic cells (DCs) from patients with chronic myeloid leukemia (CML) and attempted to elucidate its antitumor mechanisms. We isolated mononuclear cells (MNCs) from CML patients. After incubation with granulocyte macrophage colony-stimulating factor (GM-CSF), interlukin-4, and tumor necrosis factor-α (TNF-α), the MNCs developed the morphological characteristics of DCs in vitro (CML-DCs). The cells were separated according to phenotype by flow cytometric analyses with DC markers. The activity of IL- 12 and the effects of stimulation increased in CML DCs prestimulated by polysaccharide fraction of C. sinensis (PSCS) compared to untreated CML-MNCs. Furthermore, the CML-DCs incubated by PSCS resulted in the rapid generation of the costimulatory molecules, CD86 and HLA-DR, and the enhancement of IL-12 expression and stimulatory capacity in allogeneic mixed lymphocyte reaction (MLR). In summary, these results suggested that PSCS can increase T cell immunoresponse and represent a valuable traditional agent for the rapid generation of active DCs. It may also be utilized for vaccine against CML. © 2011 Academic Journals.

Shen D.,Zhejiang University | Xu C.,Yongjia Peoples Hospital | Fang H.,The First Peoples Hospital of Taizhou | Xie J.,The Second Peoples Hospital of Hangzhou | And 3 more authors.
African Journal of Biotechnology | Year: 2011

The molecular chaperone, heat shock protein gp96 (HSP-gp96), has been shown to have roles in the synthesis, processing and transport of tumor antigens. Therefore, the capacity for HSP-gp96 to induce dendritic cells (DCs), thymus-dependent lymphocytes (T lymphocytes), natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) was investigated. Recombinant adenovirus (AD) containing HSP-gp96 (AD-gp96), as well as gp96-peptide complex from the human leukemia cell lines, K562, HL-60 and U937, was prepared. Purified gp96-peptide complex was found to stimulate the proliferation of T lymphocytes, increase the activity of NK cells and CTLs and induce the secretion of cytokines, compared with ADgp96. In the latter case, levels of IFN-γ and TNF-α were found to increase and levels of IL-12(P70) and IL- 10 decreased. In combination, these results indicated that the gp96-peptide complexes derived from the tumor cells contributed to the activation of lymphocytes and increase the presentation of tumor antigen. Furthermore, the chaperone function of gp96 promoted the maturation of DCs, enhanced the antigen presention function of DCs and induced the secretion of cytokines by DCs. Therefore, gp96-peptide complex derived from the tumor cells potentially represents an immunization therapy for the elimination of residual leukemia cells. © 2011 Academic Journals.

Tong X.-M.,Zhejiang University | Zhang J.,The Center hospital of Lishui | Yuedi-Shen,The second peoples hospital of Hangzhou | Xie J.-J.,The second peoples hospital of Hangzhou | Jin J.,Zhejiang University
Journal of Medicinal Plants Research | Year: 2011

Sinomenine (SN) a component from Sinomenium acutum, widely used as an anti-inflammatory agent in Chinese traditional medicine, was reported to have anti-tumor activities. This study aims to examine the anti-leukemic activity of SN with or without aclarubicin (Acla) on leukemia cells and its molecular mechanism. HL-60 cells were treated with SN (5 to 20 ng/ml) with or without Acla (0.05 to 0.25 μg/ml). To study apoptosis, the cells were stained with Annexin V - propidium iodide (PI) and assessed by flow cytometry. To investigate the molecular mechanism, Caspase 3, Caspase 9 and cyclooxygenase-2 (Cox- 2) were assessed by Western blot, NF-kB activity and prostaglandin E2 (PGE2) were tested by ELISA. Exposure to Acla (even at 0.25 μg/ml) alone or SN alone (20 ng/ml) for 20 h did not induce significant apoptosis. However, SN (5 to 20 ng/ml) promoted apoptosis induced by 0.1 μg/ml Acla in a dosedependent manner. Increased Caspase 3 and 9 protein expression correlated positively with SN concentration when combined with 0.1 μg/ml Acla. In addition, SN significantly inhibited Acla-induced NF-kB activation, Cox-2 gene expression and PGE2 production. In summary, SN most likely enhances apoptosis by suppressing NF-kB activation, and Cox-2 and PGE2 expression. In the future, this natural agent may provide a new avenue for anti-leukemia treatment. ©2011 Academic Journals.

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