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1.
Oncol Lett ; 11(6): 3719-3722, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27313682

RESUMO

It is well known that heat shock protein 90 (HSP90) overexpression is correlated with poor prognosis and chemo-resistance in human malignant cancers. At the same time, wighteone, or 6-prenyl-5,7,4'-trihydroxyisoflavone, a major isoflavone component of the ornamental tall tree Erythrina suberosa, has been demonstrated to exhibit a potent anti-proliferative effect on human leukemia HL-60 cancer cell lines. In this study, the effects of wighteone on the proliferation of HER2-positive breast cancer cells were investigated, and the action mechanism was explored. MCF-7 HER2-positive breast cancer cells were treated with various concentrations of wighteone. The growth inhibitory rate of the cells was calculated by MTT assay, apoptosis was detected by flow cytometry, and the expression level of HSP90 was assessed by western blot analysis. The addition of wighteone at concentrations ranging from 1-10 g/ml in the medium for 48 h had a marked inhibition on the proliferation of HER2-positive cancer cell lines. The growth inhibitory rates with 0.5, 2 or 8 mM wighteone were significantly higher compared with the control group. Apoptosis in the wighteone-treated cells was also significantly higher compared with the control group. The expression level of HSP90 in the wighteone group was significantly lower than that in the control group. Our findings demonstrated that wighteone effectively inhibited the proliferation of HER2-positive cancer cell lines, and this is considered to be the result of downregulating HSP90 receptor and downstream signaling.

2.
Tumour Biol ; 34(6): 3705-12, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23873106

RESUMO

MicroRNAs (miRNAs) are important regulators of gastric cancer development and progression. miR-148a is one of the most frequently and highly downregulated miRNAs in gastric cancer and is associated with advanced clinical stage and poor prognosis. In this study, we investigated the role of miR-148a in gastric cancer metastasis. Levels of miR-148a were determined by qRT-PCR in 60 gastric cancer samples. Cell migration and invasion assays were performed in a stably expressing miRNA-148a gastric cancer cell line established using a lentivirus expression system. Epithelial-mesenchymal transition (EMT) was evaluated using qRT-PCR and Western Blots to detect epithelial marker E-cadherin and mesenchymal marker, vimentin. Luciferase reporter assays were used to identify downstream targets and biological function of miR-148a. Gastric cancer tissue had significantly lower expression of miR-148a compared to non-tumor tissue. Low miR-148a levels were associated with lymph node metastasis, N stage, and blood vessel invasion. miR-148a overexpression inhibited metastasis of gastric cancer cells. miR-148a overexpression also downregulated vimentin expression and upregulated E-cadherin expression, suggesting that miR-148a inhibited EMT. Finally, the SMAD2 gene was identified as the direct and functional target of miR-148a. MiR-148a suppresses gastric cancer metastasis and EMT, likely via SMAD2. Restoration of miR-148a expression could have important implications in gastric cancer therapy.


Assuntos
Caderinas/genética , Transição Epitelial-Mesenquimal/genética , MicroRNAs/genética , Proteína Smad2/genética , Neoplasias Gástricas/genética , Vimentina/genética , Western Blotting , Caderinas/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Regulação Neoplásica da Expressão Gênica , Células HEK293 , Humanos , Metástase Linfática , Invasividade Neoplásica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína Smad2/metabolismo , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Vimentina/metabolismo
3.
Zhonghua Nei Ke Za Zhi ; 46(9): 721-4, 2007 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-18028797

RESUMO

OBJECTIVE: To provide evidence that blocking the receptor-interacting protein 2 (Rip2) expression can decrease inflammatory cytokine production by macrophage and protect mice from endotoxin lethality. METHODS: Murine Rip2 small interfering RNA (siRNA) plamids were constructed and transfected into macrophages and Rip2 expression was assessed with RT-PCR and Western blot. Cell proliferation was assayed with MTT; TNFalpha concentration was assayed with ELISA and high-mobility group box 1 protein (HMGB1) level with semi quantitative Western blot after lipopolysaccharide (LPS) stimulation. LPS challenge was given after the plasmids were injected into mice and the survival rate was calculated. Rip2 and HMGB1 expression in liver was assessed with Western blot and serum TNFalpha level with ELISA. RESULTS: Rip2 siRNA plasmids could block the mRNA and protein expression of Rip2 and promote cell proliferation. Blocking of Rip2 could attenuate LPS-induced TNFalpha and HMGB1 production. HMGB1 expression in liver were decreased to (40.21 +/- 11.03) pg/g and serum TNFalpha level was decreased to (300.43 +/- 59.26) ng/L (P < 0.05). The survival rate of endotoxemic mice was also improved (P < 0.05). CONCLUSION: The results demonstrate that Rip2 siRNA plasmids can block the expression of Rip2, and decrease the production of TNFalpha and HMGB1, thus protect mice from lethal endotoxemia.


Assuntos
Macrófagos/metabolismo , RNA Interferente Pequeno/genética , Proteína Serina-Treonina Quinases de Interação com Receptores/genética , Animais , Western Blotting , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Proteína HMGB1/biossíntese , Lipopolissacarídeos/farmacologia , Camundongos , Camundongos Endogâmicos , Plasmídeos/genética , Interferência de RNA , Proteína Serina-Treonina Quinase 2 de Interação com Receptor , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção , Fator de Necrose Tumoral alfa/biossíntese
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