RESUMO
Screening aimed at the evaluation of the presence of small RNAs with anticancer properties in three mushrooms species, besides Boletus edulis, namely Boletus spretus (current name Baorangia emilei), Boletus pinophilus and Cantharellus cibarius, was conducted. All mushrooms yielded an ethanol insoluble and water soluble small RNA fraction purified from co-extracted polysaccharides by anion-exchange chromatography. Small RNAs from B. spretus and C. cibarius showed strong antiproliferative activity against human colon adenocarcinoma cell lines (IC50 of 5.6 µg mL-1 and 11.1 µg mL-1 for LS180 and 1.9 µg mL-1 and 12.6 µg mL-1 for HT-29 cell lines, respectively) while those isolated from B. pinophilus showed a much lower antiproliferative activity in these cells. All RNA fractions were nontoxic against CCD841 CoTr human colon epithelial cells. A detailed study of the anticancer mechanism of C. cibarius small RNAs showed that their antiproliferative activity was due to p53-dependent cell cycle arrest mediated by p21, while the proapoptotic effect was mostly dependent on the enhancement of p53 expression. Overall, small RNA fractions isolated from some edible mushrooms, namely C. cibarius, show potent antiproliferative activity without cytotoxicity to normal cells, being a potential new anticancer agent naturally present in mushrooms that we eat.
Assuntos
Agaricales/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/fisiopatologia , RNA Fúngico/farmacologia , Pequeno RNA não Traduzido/farmacologia , Antineoplásicos/isolamento & purificação , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/genética , Neoplasias do Colo/metabolismo , Células HT29 , Humanos , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , RNA Fúngico/isolamento & purificação , Pequeno RNA não Traduzido/isolamento & purificação , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismoRESUMO
Liver diseases are a growing epidemic worldwide. If unresolved, liver fibrosis develops and can lead to cirrhosis and clinical decompensation. Around 5% of cirrhotic liver diseased patients develop hepatocellular carcinoma (HCC), which in its advanced stages has limited therapeutic options and negative survival outcomes. CEPBA is a master regulator of hepatic function where its expression is known to be suppressed in many forms of liver disease including HCC. Injection of MTL-CEBPA, a small activating RNA oligonucleotide therapy (CEBPA-51) formulated in liposomal nanoparticles (NOV340- SMARTICLES) upregulates hepatic CEBPA expression. Here we show how MTL-CEBPA therapy promotes disease reversal in rodent models of cirrhosis, fibrosis, hepatosteatosis, and significantly reduces tumor burden in cirrhotic HCC. Restoration of liver function markers were observed in a carbon-tetrachloride-induced rat model of fibrosis following 2 weeks of MTL-CEBPA therapy. At 14 weeks, animals showed reduction in ascites and enhanced survival rates. MTL-CEBPA reversed changes associated with hepatosteatosis in non-alcoholic methionine and cholic-deficient diet-induced steaotic liver disease. In diethylnitrosamine induced cirrhotic HCC rats, MTL-CEBPA treatment led to a significant reduction in tumor burden. The data included here and the rapid adoption of MTL-CEBPA into a Phase 1 study may lead to new therapeutic oligonucleotides for undruggable diseases.
Assuntos
Proteínas Estimuladoras de Ligação a CCAAT/genética , Terapia Genética/métodos , Cirrose Hepática Experimental/terapia , Pequeno RNA não Traduzido/farmacologia , Ativação Transcricional , Animais , Dietilnitrosamina/toxicidade , Doença Hepática Terminal/induzido quimicamente , Doença Hepática Terminal/genética , Doença Hepática Terminal/terapia , Regulação Neoplásica da Expressão Gênica , Células Hep G2 , Humanos , Cirrose Hepática Experimental/genética , Neoplasias Hepáticas Experimentais/induzido quimicamente , Neoplasias Hepáticas Experimentais/genética , Neoplasias Hepáticas Experimentais/terapia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/etiologia , Hepatopatia Gordurosa não Alcoólica/genética , Hepatopatia Gordurosa não Alcoólica/terapia , Pequeno RNA não Traduzido/administração & dosagem , Ratos Sprague-Dawley , Ratos WistarRESUMO
Epstein-Barr virus (EBV) encoded small, non-coding, non-polyadenylated RNAs, known as EBERs are the most abundantly expressed viral transcripts in latently EBV infected cells. We found the specific role of EBERs in cell cycle progression, resistance against chemotherapeutic drug and cellular invasion in gastric cancer cells in vitro. Ectopic expression of EBERs upregulates the expression of IL-6 and activate its downstream STAT3, which is significantly involved in downregulating the expression of cell cycle inhibitor genes p21 and p27. Stable expression of EBERs regulates the activation of pFAK and pPAK1 and the expression of anti-metastatic genes RhoGDI and KAI-1 in gastric cancer cells. In addition, administration of neu-IL-6 antibody and dominant negative STAT3ß reduces chemoresistance and inhibits invasion of EBERs-expressing gastric cancer cells. Our results thus revealed a novel role of EBERs in the coordination of IL-6-STAT3 signaling pathway to chemoresistance and cellular migration.