RESUMO
The present study aimed to explore the regulatory mechanism of long intergenic nonprotein coding (LINC)00238 in hepatocellular carcinoma (HCC). LINC00238 expression in HCC tissues and cell lines was measured using reverse transcriptionquantitative PCR. LncTar was used to predict the binding sites between LINC00238 and transmembrane protein 106C (TMEM106C). Survival analysis of LINC00238, TMEM106C and activating transcription factor 3 (ATF3) in patients with HCC was performed based on TCGA data. The proliferation, apoptosis, migration, and invasion of HCC cells were measured by 3(4,5dimethylthiazol2yl)5(3carboxymethoxyphenyl)2(4sulfophenyl)2Htetrazolium assay, ï¬ow cytometer, wound healing and Transwell assays, respectively. LINC00238 promoted apoptosis and inhibited proliferation, migration and invasion of HCC cells. LINC00238 was downregulated in HCC. TMEM106C was a target of LINC00238 and TMEM106C expression was negatively regulated by LINC00238. TMEM106C suppressed the apoptosis pathway and decreased the expression of caspase7, tissue inhibitor of metalloproteinase 2, programmed cell death 4 and ATF3. Notably, ATF3 was the upstream promoter of LINC00238 and positively regulated LINC00238 expression. In conclusion, LINC00238 inhibited HCC progression by inhibiting TMEM106 expression and activating the TMEM106Cmediated apoptosis pathway.
Assuntos
Fator 3 Ativador da Transcrição/metabolismo , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Proteínas de Membrana/genética , RNA Longo não Codificante/metabolismo , Adulto , Idoso , Apoptose/genética , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/cirurgia , Linhagem Celular Tumoral , Conjuntos de Dados como Assunto , Regulação para Baixo , Retroalimentação Fisiológica , Feminino , Regulação Neoplásica da Expressão Gênica , Hepatectomia , Humanos , Fígado/patologia , Fígado/cirurgia , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/cirurgia , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Regiões Promotoras Genéticas/genética , RNA Longo não Codificante/genética , Transdução de Sinais/genéticaRESUMO
PTEN exerts tumor suppressor role through inhibiting PI3K/AKT signaling. DJ-1 plays an oncogenic role through negatively regulation of PTEN expression. Curcumin (Cur) is a phenolic compound extracted from a variety of plant roots, with multiple anti-tumor pharmacological effects. This study aims to investigate whether Cur plays a role in the regulation of DJ-1-PENT/PI3K/AKT signaling as well as the proliferation and apoptosis of hepatocellular carcinoma cells. Normal human hepatocyte HL-7702 and hepatocellular carcinoma cell lines SMMC-7721 and HepG2 were cultured followed by analysis of the expression of DJ-1 and PTEN. SMMC-7721 and HepG2 cells were treated with different concentrations of Cur (0, 5, 10 µM) followed by measuring cell proliferation by CCK-8, caspase-3 activity as well as DJ-1 expression by western blot. In addition, SMMC-7721 or HepG2 cells were divided into two groups: Cur+pcDNA3.1-Blank and Cur+pcDNA3.1-DJ-1 for analysis of the expression of DJ-1, PTEN and p-AKT, cell apoptosis and proliferation. Compared with HL-7702, SMMC-7721 and HepG2 cells displayed significantly higher DJ-1 expression and lower PTEN expression. Cur treatment significantly inhibited proliferation of SMMC-7721 and HepG2 cells, increased caspase-3 activity and downregulated DJ-1 expression. Transfection of pcDNA3.1-DJ-1 significantly increased DJ-1 and p-AKT expression, promoted cell proliferation, but decreased PTEN expression and cell apoptosis. In conclusion, Cur inhibits proliferation of hepatocellular carcinoma cells and PTEN/PI3K/AKT signaling pathway via the reduction of DJ-1 expression, which provides new insights to the anticancer effects of curcumin in hepatocellular carcinoma.
Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Curcumina/farmacologia , Neoplasias Hepáticas/tratamento farmacológico , PTEN Fosfo-Hidrolase/genética , Proteína Desglicase DJ-1/genética , Apoptose/efeitos dos fármacos , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Proliferação de Células/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND/AIMS: Liver cancer is distinguished as an irredeemable disease. We detected the geniposide (GEN) in HepG2 and Huh7 cell lines. METHODS: HepG2 and Huh7 cells were individually induced with GEN dilutions, and then they were transfected with microRNA (miR)-224 overproduction vector (miR-224 mimic) as well as the corresponding negative control (NC). Cell viability was detected with the CCK-8. The apoptotic rate was determined by the Annexin V-FITC/PI with flow cytometer. The migration or invasion rates were separately determined by migration assay or millicell hanging cell culture. The expression of miR-224 was quantified depending on qRT-PCR. Relative proteins were individually determined via western blot. RESULTS: GEN treatment induced inhibition of HepG2 and Huh7 cells proliferation, migration and invasion but promotion of apoptosis. miR-224 was down-regulated by GEN. Transfection of miR-224 mimic led to high expression of miR-224, which partly rescued cancer cells survival by prohibiting cell apoptosis. Moreover, the production of Wnt/ß-catenin and AKT proteins was notably reduced by GEN but increased by overexpressed miR-224. CONCLUSION: GEN played anti-tumor roles by targeting miR-224 via blocking the Wnt/ß-catenin and AKT cascades in the HepG2 and Huh7 cells.
