Aim: Proteins with legume lectin domains are known to possess a

Aim: Proteins with legume lectin domains are known to possess a wide range of biological functions. without altering carbohydrate structure1. Herb lectins have been divided into 12 families based on their tertiary structures and evolutionary statuses: agglutinin, Cyanovirin, Chitinase-related agglutinin, agglutinin, agglutinin (GNA), hevein, jacalin, lysin motif, proteins with legume lectin domains, nictaba, and ricin-B families. Proteins with legume lectin domains have multiple significant biological functions such as anti-fungal, anti-viral, and most notably anti-tumor activities, which have given them much attention compared with the other herb lectins2,3. Concanavalin A(ConA) is usually a long-studied representative legume lectin that reportedly diversifies human malignancy cell death by targeting programmed cell death (PCD). PCD refers to apoptosis and autophagy, which are evolutionary conversed processes for maintaining homeostasis and eliminating harmful cells4. Previous studies reported that ConA induced apoptosis in human melanoma A375 cells and murine macrophage PU5?1.8 cells. Moreover, ConA induced autophagic cell death in HeLa cells5,6,7. Therefore, ConA bears notable apoptosis- and autophagy-inducing properties, which make it a potential anti-neoplastic agent for future malignancy therapeutics. Sophora flavescens lectin (SFL) is usually a mannose-binding lectin that was isolated from Ait roots, which have been used as a traditional Chinese medicine for thousands of years. SFL is also a member of the legume lectin family Cilomilast and has been considered to be a model system in which to study the molecular basis of protein-carbohydrate interactions for several decades. Previous findings have exhibited that SFL has hemagglutinating and anti-fungal activities. Importantly, SFL can induce apoptosis in HeLa cells, thus functioning as an anti-tumor agent through a typical caspase-dependent pathway8,9,10. The mechanisms by which ConA and SFL induce malignancy cell death are only rudimentarily comprehended. In the current study, we statement that ConA and SFL Cilomilast induced apoptotic cell death in MCF-7 cells. ConA induced apoptosis via NF-B, ERK and JNK down-regulation and p53 up-regulation in human breast carcinoma MCF-7 cells. SFL reduced NF-B and ERK expression and increased p53 and p21 expression. This show that SFL initiates a G2/M phase cell-cycle arrest via up-regulating p21 and down-regulating CDK1 and CDK2 expression. Both ConA and SFL only selectively induced MCF-7 cell death but displayed no significant cytotoxicity toward normal human mammary epithelial MCF-10A cells. Furthermore, anti-tumor effects of ConA and SFL were detected, and both lectins decreased subcutaneous tumor volume and excess weight malignancy model, the cultured human breast adenocarcinoma MCF-7 cell suspension (5.0106 cells) was inoculated under the skin on the back of a 3-month-old female nude mouse. Cell proliferation and centrifugation Rabbit Polyclonal to MGST3. for 15 min, the supernatant protein content was decided using the Bio-Rad DC protein assay (Bio-Rad Laboratories, Hercules, CA, USA). Equivalent amounts of total protein were separated by 12% SDS-PAGE and transferred to nitrocellulose membranes, and the membranes were soaked in blocking buffer (5% skim milk). The following antibodies were purchased from Santa Cruz Cilomilast Biotech: caspase 3 (#sc-7148), caspase 9 (#sc-8355), cytochrome c (#sc-7159), Bax (#sc-493), Bid (#sc-6538), Bcl-2 (#sc-492), Bcl-XL (#sc-8392), NF-B (#sc-114), ERK (#sc-154), p53 (#sc-126), and -actin (#sc-47778). Antibodies including cdk1 (ab18), cdk2 (ab6538), p21 (ab7960), and JNK (ab4821) were from Abcam. Proteins were visualized using horseradish peroxidase (HRP)-conjugated anti-rabbit or anti-mouse IgG and 3,3-diaminobenzidine tetrahydrochloride (DAB) as the HRP substrate. study design In total, 40 3-month-old female nude mice were randomly divided into four groups: the blank control group (administered PBS after MCF-7 cell injection), 40 mg/kg ConA group, 55 mg/kg SFL group and the positive control group (administered cisplatin after MCF-7 cell injection). The mice were injected with PBS, ConA, SFL and cisplatin intraperitoneally, and the therapy lasted for two weeks. Animal handling was in accordance with the Ethics Committee of Sichuan University or college, and all of the animals were kept in 12-h light/dark cycles with free access to water and food, which is usually consistent with Sichuan University or college IVC requirements. Relative tumor volume, survival rate, inhibitory rate and body weight determination Tumor volume was decided from caliper measurements according to the formula, Tvol=lengthwidthdepth0.5. Tumor volume inhibitory rate=(VcontrolCVt)/Vcontrol100%. After 14 d of treatment, the mice were killed by cervical dislocation, and the subcutaneous tumor mass was decided. Tumor excess weight inhibitory rate=(WcontrolCWt)/Wcontrol100%. Statistical analysis All of the results offered here were confirmed in at least three impartial experiments. These data were expressed as the meanSEM. Statistical.

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