RESUMO
Prostatic carcinoma is the most aggressive tumor in adult men. Warburg effect is an important characteristic of tumor cell metabolism including prostate cancer cells, in which hexokinase 2 (HK2), a major rate-limiting enzyme involved in Warburg effect, is selectively upregulated. The lectin PCL is a mannose binding lectin which induces tumor cell apoptosis and autophagy. In the present study, we report that PCL could lower glucose consumption and lactate production, shift the Warburg effect by inhibiting the expression of HK2 in PC3 cells and the suppression of HK2 by siRNA reversed the effect of PCL on glucose consumption and lactate production. The expression of HK2 is closely related to epidermal growth factor receptor (EGFR) and downstream signaling pathway activation, therefore, we investigated the interaction of PCL with EGFR by western blot analysis and found that PCL could suppress the binding of epidermal growth factor (EGF) with EGFR and HK2 expression. Also, we explored the binding mechanism between the PCL and EGFR through molecular docking and molecular dynamics simulations and found that PCL bocked the active site of EGFR which is also the binding site of the nature ligand EGF, the resulting conformation has higher stability than EGF in complex with EGFR. The results indicated that PCL could competitively bind to EGFR binding pocket and then prevent EGF from binding to EGFR, blocking the autophosphorylation of the EGFR tyrosine kinase, after that the EGFR activation is inhibited. Collectively, our studies concluded that PCL inhibits tumor cell glycolysis by combining with EGFR and reducing HK2 expression.
Assuntos
Antineoplásicos/farmacologia , Receptores ErbB/metabolismo , Glicólise/efeitos dos fármacos , Hexoquinase/antagonistas & inibidores , Lectinas/farmacologia , Consumo de Oxigênio/efeitos dos fármacos , Neoplasias da Próstata/tratamento farmacológico , Adulto , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Western Blotting , Cromatografia Líquida de Alta Pressão , Ensaio de Imunoadsorção Enzimática , Receptores ErbB/química , Receptores ErbB/genética , Humanos , Masculino , Simulação de Acoplamento Molecular , Polygonatum/química , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Conformação Proteica/efeitos dos fármacos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Células Tumorais CultivadasRESUMO
AIM: To characterize the immunological function of Myostatin gene vaccine and observe the effect of Myostatin gene vaccine on the immunized animal. METHODS: The mice were immunized with Myostatin gene vaccine. We characterized the antibody titer in the immunized mice serum by the ELISA. The anti-serum was detected by auto-biochemistry analysis software. Meanwhile, the effect anti-serum was detected by auto-biochemistry analysis software. Meanwhile, the effect of Myostatin gene vaccine on skeletal-muscle of the immunized mice was analyzed by HE stain. The cross section area of muscle fiber in immunized mice was analyzed by the Scion Image 4.02 software. RESULTS: Myostatin gene vaccine could induce the anti-serum against Myostatin in immunized mice. Compared with that in the control group, the mean weight in the pVAC-TT-Ms immunized mice group increased with 9.8%. The quadriceps muscle, gastrocnemius muscle and pectoralis magor in immunized mice increased with 24.1%, 10.9% and 20.3%, respectively. CONCLUSION: Myostatin gene vaccine induced the specific antibody against Myostatin, and made the muscle strong.
