RESUMO
The focus of the study is to examine the function of TYMSOS in immune escape of breast cancer, which is the most frequently diagnosed malignancy among women globally. Our study demonstrated that upregulated TYMSOS was associated with unfavorable prognosis and immune escape in breast cancer. TYMSOS promoted the malignant phenotypes of breast cancer cells, and reduced the cytotoxicity of NK92 cells on these cells. CBX3 was a downstream effector in TYMSOS-induced malignant phenotypes in breast cancer cells. Mechanistic studies showed that TYMSOS facilitated CBX3-mediated transcriptional repression of ULBP3, and it also promoted SYVN1-mediated ubiquitin-proteasomal degradation of ULBP3. TYMSOS promoted cell growth, metastasis, and immune escape via CBX3/ULBP3 or SYVN1/ULBP3 axis. The in vivo studies further showed that silencing of TYMSOS repressed tumor growth and boosted NK cell cytotoxicity. In sum, TYMSOS boosted breast cancer metastasis and immune escape via CBX3/ULBP3 or SYVN1/ULBP3 axis.
RESUMO
Long non-coding RNA (lncRNA) is involved in the regulation of tumorigenesis and metastasis. In this study, we focused on the clinical relevance, biological effects, and molecular mechanisms of the lncRNA differentiation antagonizing non-protein coding RNA (DANCR) in breast cancer. We compared the expression of DANCR between breast cancer and normal tissues, and between breast cancer cell lines and normal breast epithelial cells using quantitative real-time PCR (qRT-PCR) analysis. By knocking down and overexpressing DANCR, we assessed its significance in regulating viability (MTT assay), migration/invasion (Transwell assay), epithelial-mesenchymal transition (western blot), stemness (mammosphere formation assay and western blot), and production of inflammatory cytokines (qRT-PCR and ELISA) of breast cancer cells in vitro, as well as xenograft growth in vivo. Furthermore, using ChIP and RNA immunoprecipitation, we examined the reciprocal regulation between DANCR and suppressor of cytokine signaling 3 (SOCS3) in breast cancer. DANCR was significantly up-regulated in tissue samples from patients with breast cancer, as well as in breast cancer cell lines, as compared with normal tissues and breast epithelial cells, respectively. The highest DANCR expression levels were associated with advanced tumor grades or lymph node metastasis. DANCR was necessary and sufficient to control multiple malignant phenotypes of breast cancer cells in vitro and xenograft growth in vivo. Mechanistically, DANCR promoted the binding of enhancer of zeste homolog 2 (EZH2) to the promoter of SOCS3, thereby epigenetically inhibiting SOCS3 expression. Functionally, SOCS3 up-regulation or EZH2 inhibition could rescue multiple malignant phenotypes induced by DANCR. Our data indicate that DANCR is a pleiotropic oncogenic lncRNA in breast cancer. Boosting SOCS3 expression may reverse the oncogenic activities of DANCR and thus provide a therapeutic strategy for breast cancer treatment.
Assuntos
Neoplasias da Mama/metabolismo , Citocinas/metabolismo , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , RNA Longo não Codificante/metabolismo , Proteína 3 Supressora da Sinalização de Citocinas/metabolismo , Animais , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Movimento Celular/genética , Proliferação de Células/genética , Sobrevivência Celular/genética , Imunoprecipitação da Cromatina , Progressão da Doença , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Transição Epitelial-Mesenquimal/genética , Feminino , Humanos , Inflamação/metabolismo , Metástase Linfática/genética , Células MCF-7 , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Regiões Promotoras Genéticas , Ligação Proteica , RNA Longo não Codificante/genética , Proteína 3 Supressora da Sinalização de Citocinas/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Previously we showed that aldehyde dehydrogenase 1A1 (ALDH1A1) is a new mediator for resistance of DLBCL to CHOP and a facility predictor of clinical prognosis. In the present study, knockdown and inhibitor of ALDH1A1 were applied to identify the role of ALDH1A1 in Raji cells. CCK-8 and clone formation assay were applied to determine the CHOP sensitivity and clone formation ability. Caspase colorimetric assay and Annexin V/FITC staining was performed to determine the degree of apoptosis. Western blot analysis was used to detect the NF-κB/STAT3 signaling proteins and apoptotic-associated proteins. Real-time quantitative PCR (RT-PCR) was used to identify the differential expression of ALDH1A1 between NHL patients and healthy donors. We demonstrated that inhibition of ALDH1A1 increased the sensitivity of Raji cells to CHOP, as indicated by increased cytotoxicity, reduced clonogenicity, activated caspase-3/-9, decreased NF-κB/STAT3 signaling and increased pro-apoptosis signaling, ad increased apoptosis rate. Moreover, we found high ALDH1A1 expression was associated with poor prognosis in NHL patients. Our data revealed the critical role of ALDH1A1 in NHL and provides a theoretical basis for the use of ALDH1A1 inhibitors in NHL patients.
Assuntos
Aldeído Desidrogenase/fisiologia , Linfoma de Células B/enzimologia , Aldeído Desidrogenase/antagonistas & inibidores , Aldeído Desidrogenase/metabolismo , Família Aldeído Desidrogenase 1 , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Apoptose , Linhagem Celular Tumoral , Ciclofosfamida/farmacologia , Doxorrubicina/farmacologia , Humanos , Linfoma de Células B/mortalidade , NF-kappa B/metabolismo , Prednisona/farmacologia , Prognóstico , Retinal Desidrogenase , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais , Vincristina/farmacologiaRESUMO
Breast cancer (BC) is the foremost cause of cancer-related mortality in women worldwide. Polyporus umbellatus is a polysaccharide preparation of the Chinese traditional herb medicine, which has been explored as an inhibitory compounds in suppressing many cancers. And AKT has been known as an essential signaling pathway to regulate cell proliferation and apoptosis via Mdm2/p53 and Caspase-3 signaling pathways respectively. In our study, western blot, RT-PCR, immunochemical assay, immunofluorescence as well as flow cytometry were performed in vitro or in vivo to determine the effects of Polyporus umbellatus on the progression of human laryngeal cancer. First, the breast cancer cell growth, invasion and migration were inhibited, as well as the tumor volume in nude mice was down-regulated for Polyporus umbellatus use. Additionally, our data also showed that Polyporus umbellatus suppressed breast cancer cells proliferation, which was linked with the down-regulation of AKT activation by Polyporus umbellatus treatment. Mdm was inactivated while p53 was stimulated for Polyporus umbellatus administration, displaying inhibitory role in tumor growth. Furthermore, Polyporus umbellatus could up-regulate breast cancer cells in G0/G1 phase during cell cycle, and at the same time reducing cells in S phase. Also, flow cytometry and western blot assays suggested that apoptosis was induced by the administration of Polyporus umbellatus, which enhanced Caspase-3 expressions by AKT-regulated anti-apoptotic and pro-apoptotic signals. In conclusion, our data indicated that Polyporus umbellatus had a potential role in controlling human breast cancer through inhibiting tumor cell proliferation, inducing apoptosis regulated by AKT, which might provide a therapeutic strategy for breast cancer suppression in the future.