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1.
FASEB J ; 35(9): e21332, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34423867

RESUMO

Emerging research has highlighted the capacity of microRNA-23a-3p (miR-23a-3p) to alleviate inflammatory pain. However, the molecular mechanism by which miR-23a-3p attenuates inflammatory pain is yet to be fully understood. Hence, the current study aimed to elucidate the mechanism by which miR-23a-3p influences inflammatory pain. Bioinformatics was initially performed to predict the inflammatory pain related downstream targets of miR-23a-3p in macrophage-derived extracellular vesicles (EVs). An animal inflammatory pain model was established using Complete Freund's Adjuvant (CFA). The miR-23a-3p expression was downregulated in the microglia of CFA-induced mice, after which the inflammatory factors were determined by ELISA. FISH and immunofluorescence were performed to analyze the co-localization of miR-23a-3p and microglia. Interestingly, miR-23a-3p was transported to the microglia via M2 macrophage-EVs, which elevated the mechanical allodynia and the thermal hyperalgesia thresholds in mice model. The miR-23a-3p downstream target, USP5, was found to stabilize HDAC2 via deubiquitination to promote its expression while inhibiting the expression of NRF2. Taken together, the key findings of the current study demonstrate that macrophage-derived EVs containing miR-23a-3p regulates the HDAC2/NRF2 axis by decreasing USP5 expression to alleviate inflammatory pain, which may provide novel therapeutic targets for the treatment of inflammatory pain.


Assuntos
Vesículas Extracelulares/metabolismo , Histona Desacetilase 2/metabolismo , Inflamação/metabolismo , Macrófagos/citologia , Fator 2 Relacionado a NF-E2/metabolismo , Dor/metabolismo , Proteases Específicas de Ubiquitina/metabolismo , Animais , Linhagem Celular , Enzimas Desubiquitinantes/metabolismo , Modelos Animais de Doenças , Regulação para Baixo , Estabilidade Enzimática , Vesículas Extracelulares/genética , Inflamação/genética , Inflamação/terapia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microglia/citologia , Microglia/metabolismo , Modelos Biológicos , Dor/genética , Manejo da Dor , Proteases Específicas de Ubiquitina/genética , Ubiquitinação
2.
Int J Mol Med ; 47(1): 161-170, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33416107

RESUMO

The mitochondria have been proven to be involved in processes of aging; however, the mechansims through which mitoepigenetics affect the cytological behaviors of cardiomyocytes during the aging process are not yet fully understood. In the present study, two senescence models were constructed, replicative senescence (RS) and stress­induced premature senescence (SIPS), using human heart mesenchymal stem cells (HMSCs). First, the differences in age­related gene expression levels and telomere length were compared between the HMSCs in the RS and SIPS models by PCR. Subsequently, protein expression and the mitochondrial DNA (mtDNA) methylation status of cytochrome c oxidase subunit II (COX2) was measured by western blot analysis and bisulfite genomic sequencing (BSP). Finally, the value of the DNA methyltransferase (Dnmt) inhibitor, 5­aza­2'­deoxycytidine (AdC), in delaying the senescence of HMSCs was evaluated. It was found that the p16, p27 and p53 mRNA expression levels increased in the senescent cells, whereas p21 mRNA expression did not. It was also found that telomere shortening only occurred in the RS model, but not in the SIPS model. Along with the senescence of HMSCs, COX2 gene methylation increased and its protein expression level significantly decreased. It was demonstrated that AdC inhibited COX2 methylation and downregulated COX2 expression. The addition of exogenous COX2 or the administration of AdC promoted cell proliferation and delayed cell aging. On the whole, the present study demonstrates that COX2 methylation and downregulation are biomarkers of HMSC senescence. Thus, COX2 may have potential for use as a therapeutic target of cardiovascular diseases and this warrants further investigation.


Assuntos
Senescência Celular , Metilação de DNA , DNA Mitocondrial/metabolismo , Regulação para Baixo , Complexo IV da Cadeia de Transporte de Elétrons/biossíntese , Regulação Enzimológica da Expressão Gênica , Células-Tronco Mesenquimais/enzimologia , Mitocôndrias Cardíacas/enzimologia , Miocárdio/enzimologia , DNA Mitocondrial/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Humanos , Mitocôndrias Cardíacas/genética
3.
Ying Yong Sheng Tai Xue Bao ; 31(7): 2373-2380, 2020 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-32715703

RESUMO

Application of organic amendments is an effective approach for improving soil organic carbon and soil fertility. To investigate the effects of different organic amendments on soil organic carbon and its labile fraction content, a batch of incubation experiments was conducted on the fluvo-aquic soil in Dongting Lake region, Hunan Province. There were six treatments, including soil amended with rice straw, soil amended with Chinese milk vetch, soil amended with bio-organic fertilizer, soil amended with pig manure, and soil amended with rice straw-derived biochar, with unamended soil as control. Each treatment had the same amount of carbon input. After 180 days of incubation, application of organic amendments increased soil labile organic carbon content. Application of bio-organic fertilizer, pig manure and rice straw-derived biochar significantly increased soil organic carbon content by 26.1%, 9.7% and 30.7%, respectively. There was no significant change in soil organic carbon content in rice straw and Chinese milk vetch treatments which were more favourable to the accumulation of soil dissolved organic carbon and microbial biomass carbon. Pig manure was more favourable to the accumulation of soil dissolved organic carbon. Bio-organic fertili-zer could benefit the accumulation of soil microbial biomass carbon and readily oxidizable organic carbon. Rice straw-derived biochar could promote the accumulation of soil microbial biomass carbon and light fraction organic carbon. Compared with rice straw, soil carbon pool management index was increased by 31.8%, 111.6%, 62.2% and 50.7% in Chinese milk vetch, bio-organic fertilizer, pig manure and rice straw-derived biochar treatments, respectively. The performance of bio-organic fertilizer, pig manure, and rice straw biochar was better than rice straw and Chinese milk vetch from the perspective of soil carbon sequestration and soil carbon pool management index.


Assuntos
Oryza , Solo , Agricultura , Animais , Carbono , Carvão Vegetal , Fertilizantes , Suínos
4.
Adv Sci (Weinh) ; 7(12): 2000177, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32596119

RESUMO

It is remarkably desirable and challenging to design a stretchable conductive material with tunable electromagnetic-interference (EMI) shielding and heat transfer for applications in flexible electronics. However, the existing materials sustained a severe attenuation of performances when largely stretched. Here, a super-stretchable (800% strain) liquid metal foamed elastomer composite (LMF-EC) is reported, achieving super-high electrical (≈104 S cm-1) and thermal (17.6 W mK-1) conductivities under a large strain of 400%, which also exhibits unexpected stretching-enhanced EMI shielding effectiveness of 85 dB due to the conductive network elongation and reorientation. By varying the liquid and solid states of LMF, the stretching can enable a multifunctional reversible switch that simultaneously regulates the thermal, electrical, and electromagnetic wave transport. Novel flexible temperature control and a thermoelectric system based on LMF-EC is furthermore developed. This work is a significant step toward the development of smart electromagnetic and thermal regulator for stretchable electronics.

5.
Adv Mater ; 32(17): e2000827, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32134520

RESUMO

It is remarkably desirable and challenging to design reconfigurable ferromagnetic materials with high electrical conductivity. This has attracted great attention due to promising applications in many fields such as emerging flexible electronics and soft robotics. However, the shape and magnetic polarity of existing ferromagnetic materials with low conductivity are both hard to be reconfigured, and the magnetization of insulative ferrofluids is easily lost once the external magnetic field is removed. A novel reconfigurable ferromagnetic liquid metal (LM) putty-like material (FM-LMP) with high electrical conductivity and transformed shape, which is prepared through homogenously mixing neodymium-iron-boron microparticles into the gallium-based LM matrix, and turning this liquid-like suspension into the solid-like putty-like material by magnetization, is reported to achieve this. The induction magnetic field of FM-LMP is mainly attributed to the magnetic alignment of the dispersed ferromagnetic microparticles, which can be conveniently demagnetized by mechanical disordering and reversibly reconfigured through microparticle realignment by applying a weak magnetic field. FM-LMP with a low fraction of microparticles can be used as printable conductive ink for paper electronics, which are further exploited for applications including magnetic switching, flexible erasable magnetic recording paper, and self-sensing paper-based soft robotics using magnetic actuation.

6.
Oncol Rep ; 43(3): 897-907, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32020207

RESUMO

Radioresistance hinders the therapeutic outcomes of radiotherapy in non­small cell lung cancer (NSCLC). Although long non­coding RNAs (lncRNAs) have been demonstrated to participate in the regulation of multiple cell behaviors, whether they can modulate the radiosensitivity of NSCLC and the underlying molecular mechanisms have not been well investigated. In the present study, it was revealed that NSCLC NCI­H460 cells were more sensitive to ionizing radiation (IR) than A549 cells. Using the RNA­Seq method, four highly differentially expressed lncRNAs were identified, including the growth arrest­specific transcript 5 (GAS5), syntaxin binding protein 5 antisense RNA 1 (STXBP5­AS1), metastasis associated lung adenocarcinoma transcript 1 (MALAT1) and X­inactive specific transcript (XIST), which were predicted to play roles in the acquisition of radiosensitivity. Using real­time quantitative PCR (qPCR), it was demonstrated that lncRNA GAS5 was significantly upregulated in NCI­H460 cells but not in A549 cells during IR. Mechanistically, it was demonstrated that overexpression of lncRNA GAS5 decreased the level of microRNA­21 (miR­21). Overexpression of lncRNA GAS5 or suppression of miR­21 markedly increased the IR­induced cell apoptosis of A549 cells. It was also demonstrated that overexpression of lncRNA GAS5 increased PTEN expression and suppressed Akt phosphorylation through the modulation of miR­21. Notably, it was revealed that IR enhanced the interaction between lncRNA GAS5 and the miR­21/PTEN/Akt axis. In summary, the present findings revealed that lncRNA GAS5 has a radiosensitization effect on NSCLC, indicating the potential application of lncRNA GAS5 in NSCLC radiotherapy.


Assuntos
Neoplasias Pulmonares/genética , MicroRNAs/genética , PTEN Fosfo-Hidrolase/genética , RNA Longo não Codificante/genética , Células A549 , Apoptose/genética , Sequência de Bases/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/radioterapia , Proteína Oncogênica v-akt/genética , Fosforilação/genética , Tolerância a Radiação/genética , Radioterapia
7.
Carbohydr Polym ; 230: 115580, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31887929

RESUMO

It is a significant challenge to reduce the sheet formation time and simultaneously sustain high transparency and good formation of nanostructured paper made of cellulose fibrils. In this study, a new strategy was developed for expediting the filtration but avoiding the flocculation of cellulose fibrils by using polyvinylpyrrolidone (PVP) and Laponite as dispersants to reduce the viscosity of the cellulose fibril aqueous dispersion. By adding 0.5 % PVP and 0.2 % Laponite to the cellulose fibril dispersion, the sheet formation time was reduced from 135.3-53.0 min, and the transmittance of the nanostructured paper was increased from 82.8%-89.8 %. The addition of PVP and Laponite significantly reduced the roughness and density but improved the tensile strength of the nanostructured paper. This study provides an approach for simultaneously improving the drainability, transparency, tensile strength, and smoothness of cellulose nanostructured paper made of cellulose fibrils.

8.
Int J Biol Macromol ; 145: 944-949, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31669275

RESUMO

Polysaccharide has been considered as an important bioactive compound in Codonopsis lanceolata. High fat/high sucrose (HFHS) diet-induced insulin resistance is implicated in multiple metabolic diseases, such as type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD), these metabolic diseases has become epidemic health issue worldwide. In this study, the effect of C. lanceolata polysaccharide (CLPS) on improving insulin sensitivity in chronic HFHS diet-fed mice was investigated. Our data indicates that CLPS significantly reduced fasting blood glucose (FBG), fasting serum insulin (FINS) and insulin resistance index, in parallel with improved glucose and insulin tolerance impaired by HFHS diet. Impaired phosphorylation of PKB/Akt and hyperphosphorylation of IRS-1 at Ser307 were observed in the mice fed with HFHS diet, and those defects were also rescued by CLPS administration. In addition, CLPS caused a significant decrease in the level of malondialdehyde (MDA), and an increase in reduced glutathione (GSH)/oxidised glutathione (GSSG) ratio; concurrent with enhanced expression of antioxidant enzymes including superoxide dismutase (SOD) and catalase (CAT), and activated Nrf2 signaling. In summary, these findings suggest that CLPS ameliorates HFHS diet-induced insulin resistance through activating anti-oxidative signaling pathway, providing new insights into the protective effects of C. lanceolata polysaccharide in metabolic disease.


Assuntos
Codonopsis/química , Dieta da Carga de Carboidratos/efeitos adversos , Dieta Hiperlipídica/efeitos adversos , Resistência à Insulina/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Polissacarídeos/farmacologia , Animais , Antioxidantes , Glicemia/efeitos dos fármacos , Catalase , China , Diabetes Mellitus Tipo 2/metabolismo , Glucose/metabolismo , Glutationa , Insulina/sangue , Insulina/metabolismo , Masculino , Malondialdeído , Camundongos , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/metabolismo , Hepatopatia Gordurosa não Alcoólica/metabolismo , Fosforilação , Raízes de Plantas/química , Polissacarídeos/química , Transdução de Sinais/efeitos dos fármacos , Superóxido Dismutase
9.
J Breast Cancer ; 22(2): 237-247, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31281726

RESUMO

Purpose: Breast cancer (BC) is one of the most common malignancies globally, and millions of women worldwide are diagnosed with BC every year. Up to 70% of BC patients are estrogen receptor (ER)-positive. Numerous studies have shown that tamoxifen has a significant therapeutic effect on both primary and metastatic ER-positive BC patients. Although tamoxifen is currently one of the most successful therapeutic agents for BC, a significant proportion of patients will eventually become resistant to tamoxifen, leading to tumor recurrence and metastasis. Knowledge about the development of tamoxifen resistance in BC patients is still limited. Methods: We applied a loss-and-gain method to study the biological functional role of cell division cycle associated 8 (CDCA8) in tamoxifen resistance in BC cells. Results: We found that CDCA8 was significantly elevated in tamoxifen-resistant BC cells. Knockdown of CDCA8 expression significantly inhibited the proliferation of tamoxifen-resistant BC cells and reduced their resistance to tamoxifen. In contrast, overexpression of CDCA8 promoted the growth of tamoxifen-sensitive BC cells and induced their resistance to tamoxifen. Conclusion: In this study, we reported that CDCA8 is a key regulator of tamoxifen resistance in BC, suggesting that CDCA8 may serve as a potential therapeutic target for BC treatment.

10.
Genome Res ; 29(9): 1521-1532, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31315906

RESUMO

Long noncoding RNAs (lncRNAs) can regulate the activity of target genes by participating in the organization of chromatin architecture. We have devised a "chromatin-RNA in situ reverse transcription sequencing" (CRIST-seq) approach to profile the lncRNA interaction network in gene regulatory elements by combining the simplicity of RNA biotin labeling with the specificity of the CRISPR/Cas9 system. Using gene-specific gRNAs, we describe a pluripotency-specific lncRNA interacting network in the promoters of Sox2 and Pou5f1, two critical stem cell factors that are required for the maintenance of pluripotency. The promoter-interacting lncRNAs were specifically activated during reprogramming into pluripotency. Knockdown of these lncRNAs caused the stem cells to exit from pluripotency. In contrast, overexpression of the pluripotency-associated lncRNA activated the promoters of core stem cell factor genes and enhanced fibroblast reprogramming into pluripotency. These CRIST-seq data suggest that the Sox2 and Pou5f1 promoters are organized within a unique lncRNA interaction network that determines the fate of pluripotency during reprogramming. This CRIST approach may be broadly used to map lncRNA interaction networks at target loci across the genome.


Assuntos
Cromatina/genética , Fator 3 de Transcrição de Octâmero/genética , RNA Longo não Codificante/genética , Fatores de Transcrição SOXB1/genética , Análise de Sequência de RNA/métodos , Animais , Sistemas CRISPR-Cas , Linhagem Celular , Reprogramação Celular , Fibroblastos/citologia , Fibroblastos/metabolismo , Camundongos , Células-Tronco Pluripotentes/citologia , Células-Tronco Pluripotentes/metabolismo , Regiões Promotoras Genéticas , Sequências Reguladoras de Ácido Nucleico
11.
Gene ; 703: 1-6, 2019 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-30953709

RESUMO

Endocrine therapy is effective in the early stage of breast cancer treatment, and most tumor cells will gain the ability to proliferate under residual amounts of estrogen, which will cause the recurrence of the disease. The role of cell division cycle associated 8 (CDCA8) in Estradiol (E2)-stimulated breast cancer cells growth is investigated in this research. CDCA8 showed higher mRNA expression in E2-stimulated MCF7 and T47D cells, and such an increase could also be observed in tumor samples. CDCA8 shRNA inhibited the survival and growth detected by cell number and colony formation, while promoted cell cycle G1 phase arrest determined with flow cytometry, which coordinated with a decrease in E2-induced molecules, namely Cyclin D1 (CCND1), B-Cell CLL/Lymphoma 2 (BCL2), and an increase in apoptosis-related molecules, such as cyclin-dependent kinase inhibitor 1a (P21) and cyclin-dependent kinase inhibitor 1b (P27). Kaplan-Meier plot analysis indicated that higher CDCA8 expression was positively associated with poor prognosis with a probability lower than 0.4 at the five-year interval (p = 0.035). All of these suggest that CDCA8 is a key mediator of estrogen-stimulated breast cancer cell growth and survival, which can be utilized as a novel target in breast cancer treatment.


Assuntos
Neoplasias da Mama/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Estradiol/farmacologia , Regulação para Cima , Neoplasias da Mama/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Estimativa de Kaplan-Meier , Células MCF-7 , Prognóstico , RNA Interferente Pequeno/farmacologia , Regulação para Cima/efeitos dos fármacos
12.
Carbohydr Polym ; 214: 26-33, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30925996

RESUMO

Fabricating transparent paper from cellulose nanofibers (CNFs) normally involves high energy or the use of expensive chemicals for the extraction of CNFs from cellulose fibers and time-consuming paper formation processes because of the slow filtration rate of CNFs. In this study, we reported a strategy for the fabrication of transparent paper using microfibrillated cellulose fibers (MFCFs), which were prepared by extracting nanosized fibrils from the cellulose fiber surfaces by a two-step refining process. The paper made from MFCFs has hierarchical structures of microsized fiber/nanosized fibril networks, where the microsized fiber skeletons are buried in the nanosized fibril networks. Consequently, the paper shows a light transmittance of 82.4% at 550 nm; this is comparable to the light transmittance (89.1%) of papers made from CNFs. Meanwhile, the filtration time of the paper made from MFCFs is less than 2 min, which is much shorter than the time (longer than 180 min) required for the formation of nanopaper made from commercial CNFs. In addition, the transparent paper made from MFCFs shows higher thermal stability, higher tensile strength, higher resistance to deformation, and more flexibility than the nanopaper made from commercial CNFs. This work provides a promising method for the manufacture of transparent paper from cellulose fibers.


Assuntos
Celulose/química , Nanofibras/química , Papel , Cristalização , Fenômenos Ópticos , Maleabilidade , Porosidade , Resistência à Tração
13.
Int J Oncol ; 54(3): 1033-1042, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30628639

RESUMO

Tamoxifen is the gold standard for breast cancer endocrinotherapy. However, drug resistance remains a major limiting factor of tamoxifen treatment. Long non­coding (lnc) RNA serves an important role in drug resistance; however, the molecular mechanisms of tamoxifen resistance in breast cancer endocrinotherapy are largely unclear. lncRNA urothelial cancer associated 1 (lncRNA UCA1, UCA1) has been proven to be dysregulated in human breast cancer and promotes cancer progression. In the present study, it was demonstrated that UCA1 was significantly upregulated in breast cancer tissues compared with healthy tissues. Furthermore, the expression level of UCA1 was significantly greater in tamoxifen­resistant breast cancer cells (LCC2 and LCC9) when compared with those in the tamoxifen­sensitive breast cancer cells (MCF­7 and T47D). UCA1 silencing in LLC2 and LLC9 cells increased tamoxifen drug sensitivity by promoting cell apoptosis and arresting the cell cycle at the G2/M phase. Notably, the induced overexpression of UCA1 in MCF­7 and T47D cells decreased the drug sensitivity of tamoxifen. The molecular mechanism involved in UCA1­induced tamoxifen­resistance was also investigated. It was identified that UCA1 was physically associated with the enhancer of zeste homolog 2 (EZH2), which suppressed the expression of p21 through histone methylation (H3K27me3) on the p21 promoter. In addition, it was demonstrated that UCA1 expression was paralleled to the phosphorylation of CAMP responsive element binding protein (CREB) and AKT. When LCC2 cells were treated with the phosphoinositide 3­kinase (PI3K)/protein kinase B (AKT) signaling pathway inhibitor LY294002, the phosphorylation levels of CREB and AKT were significantly downregulated. Taken together, it was concluded that UCA1 regulates the EZH2/p21 axis and the PI3K/AKT signaling pathway in breast cancer, and may be a potential therapeutic target for solving tamoxifen resistance.


Assuntos
Antineoplásicos Hormonais/farmacologia , Neoplasias da Mama/genética , Resistencia a Medicamentos Antineoplásicos/genética , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , RNA Longo não Codificante/metabolismo , Transdução de Sinais , Tamoxifeno/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Ciclo Celular , Linhagem Celular Tumoral , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Inibidor de Quinase Dependente de Ciclina p21/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Longo não Codificante/genética
14.
Oncol Lett ; 17(1): 630-637, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30655810

RESUMO

In the present study, the functions and mechanisms of rotundic acid (RA) underlying its induction of apoptosis in caspase-3-transfected MCF-7 human breast cancer cells (Cas3-MCF-7 cells) were investigated. RA induced apoptosis in Cas3-MCF-7 cells more efficiently compared with that in MCF-7 cells transfected with control plasmid. The results from an MTT assay demonstrated that RA effectively inhibited Cas3-MCF-7 cell viability in a dose-dependent manner and induced cell apoptosis via caspase-3 activity within 12 to 48 h. Western blotting and fluorescence-activated cell sorting demonstrated that RA initiated Cas3-MCF-7 cell apoptosis via p53 activation. The silencing of the p53 gene in the Cas3-MCF-7 cell line led to decreased RA-induced Cas3-MCF-7 cell caspase-3 activity and cell apoptosis. Collectively, the results of the present study indicate that caspase-3 serves a critical function in rotundic acid-induced apoptosis, and suggest that caspase-3 deficiency may contribute to the chemotherapy-resistance of breast cancer. Reconstitution of caspase-3 sensitizes MCF-7 breast cancer cells to chemotherapy. RA has the potential for development as a novel drug combined with reconstitution of caspase-3 gene therapy for the treatment of human breast cancer with caspase-3 deficiency.

15.
Clin Cancer Res ; 25(4): 1302-1317, 2019 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-30429198

RESUMO

PURPOSE: The aberrantly upregulated Friend leukemia virus integration 1 (FLI1) is closely correlated with the malignant phenotype of small cell lung cancer (SCLC). It is interesting to note that the CRISPR gene knockout by Cas9 gRNAs that target the FLI1 coding region and the posttranscriptional knockdown by shRNAs that target the 3' region of FLI1 mRNA yielded distinct antimetastasis effects in SCLC cells. This study attempts to examine if FLI1 exonic circular RNAs (FECR) function as a new malignant driver that determines the metastatic phenotype in SCLC. EXPERIMENTAL DESIGN: The clinical relevance of FECRs was examined in 56 primary SCLC tissues and 50 non-small cell lung cancer (NSCLC) tissues. The prognostic value of FECRs was examined by measuring serum exosomal FECRs in a longitudinal cohort of patients with SCLC. The oncogenic activity of FECRs was investigated in both SCLC cell lines and animal xenograft studies. Finally, we explored the molecular mechanisms underlying these noncoding RNAs as a malignant driver. RESULTS: Therapeutic comparison of CRISPR Cas9 knockout and shRNA knockdown of FLI1 identified FECRs as a new noncanonical malignant driver in SCLC. Using RNA FISH and quantitative PCR, we found that FECR1 (exons 4-2-3) and FECR2 (exons 5-2-3-4) were aberrantly upregulated in SCLC tissues (P < 0.0001), and was positively associated with lymph node metastasis (P < 0.01). Notably, serum exosomal FECR1 was associated with poor survival (P = 0.038) and clinical response to chemotherapy. Silencing of FECRs significantly inhibited the migration in two highly aggressive SCLC cell lines and reduced tumor metastasis in vivo. Mechanistically, we uncovered that FECRs sequestered and subsequently inactivated tumor suppressor miR584-3p, leading to the activation of the Rho Associated Coiled-Coil Containing Protein Kinase 1 gene (ROCK1). CONCLUSIONS: This study identifies FLI1 exonic circular RNAs as a new oncogenic driver that promotes tumor metastasis through the miR584-ROCK1 pathway. Importantly, serum exosomal FECR1 may serve as a promising biomarker to track disease progression of SCLC.


Assuntos
MicroRNAs/genética , Proteína Proto-Oncogênica c-fli-1/genética , RNA Circular/genética , Carcinoma de Pequenas Células do Pulmão/genética , Quinases Associadas a rho/genética , Células A549 , Animais , Apoptose/genética , Sistemas CRISPR-Cas/genética , Carcinoma Pulmonar de Células não Pequenas/sangue , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Ciclo Celular/genética , Movimento Celular/genética , Proliferação de Células/genética , Éxons/genética , Exossomos/genética , Xenoenxertos , Humanos , Camundongos , Metástase Neoplásica , Proteína Proto-Oncogênica c-fli-1/antagonistas & inibidores , RNA Circular/isolamento & purificação , RNA Interferente Pequeno/genética , Carcinoma de Pequenas Células do Pulmão/sangue , Carcinoma de Pequenas Células do Pulmão/patologia
16.
Sci Data ; 5: 180255, 2018 11 20.
Artigo em Inglês | MEDLINE | ID: mdl-30457566

RESUMO

Pluripotent stem cells hold great investigative potential for developmental biology and regenerative medicine. Recent studies suggest that long noncoding RNAs (lncRNAs) may function as key regulators of the maintenance and the lineage differentiation of stem cells. However, the underlying mechanisms by which lncRNAs affect the reprogramming process of somatic cells into pluripotent cells remain largely unknown. Using fibroblasts and induced pluripotent stem cells (iPSCs) at different stages of reprogramming, we performed RNA transcriptome sequencing (RNA-Seq) to identify lncRNAs that are differentially-expressed in association with pluripotency. An RNA reverse transcription-associated trap sequencing (RAT-seq) approach was then utilized to generate a database to map the regulatory element network for lncRNA candidates. Integration of these datasets can facilitate the identification of functional lncRNAs that are associated with reprogramming. Identification of lncRNAs that regulate pluripotency may lead to new strategies for enhancing iPSC induction in regenerative medicine.


Assuntos
Reprogramação Celular/genética , Células-Tronco Pluripotentes , RNA Longo não Codificante , Transcriptoma , Diferenciação Celular/genética , Fibroblastos/citologia , Fibroblastos/metabolismo , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Células-Tronco Pluripotentes/citologia , Células-Tronco Pluripotentes/metabolismo , RNA Longo não Codificante/análise , RNA Longo não Codificante/genética , Análise de Sequência de RNA
17.
Int J Oncol ; 53(5): 2269-2277, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30226600

RESUMO

Although radiation therapy is a powerful anticancer modality, radiation- induced stress response and gene expression with adaptive resistance may severely compromise the effectiveness of radiation. The function of rotundic acid (RA) on inducing apoptosis in the human breast cancer cell line MCF-7 has been investigated in a previous study. In the present study, the combined effect of chemotherapy and radiotherapy on reducing side effects was examined. The results of an MTT assay revealed that radiation (0.5, 2 and 10 Gy) effectively inhibit MCF-7 cell viability in a dose-dependent manner, consistent with the effects of RA (2, 5 and 12.5 µM). Interestingly, a lower dose of radiation (1 Gy) combined with RA (5 µM) exhibited a greater inhibition efficiency compared with a high dose of radiation alone. Flow cytometry revealed that radiation combined with RA induced the apoptosis of MCF-7 cells. Using western blotting, it was demonstrated that radiation induced the expression of ataxia-telangiectasia mutated (ATM) and p53 protein, and that RA enhanced this effect. On examining the potential underlying mechanism, it was revealed that radiation and RA combined induce Bcl-2-associated X protein expression and cell apoptosis in MCF-7 cells. An ATM inhibitor was able to restore the effect of radiation and RA on inducing MCF-7 cell apoptosis. These results suggest that the ATM/p53 pathway directly participates in radiation and RA-induced apoptosis in MCF-7 cells. RA has the potential for development as a novel drug for the treatment of human breast cancer combined with radiation therapy, given that the combined side effects are reduced.


Assuntos
Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Neoplasias da Mama/terapia , Tolerância a Radiação/efeitos dos fármacos , Triterpenos/farmacologia , Proteína Supressora de Tumor p53/metabolismo , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Neoplasias da Mama/patologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Quimiorradioterapia/efeitos adversos , Quimiorradioterapia/métodos , Relação Dose-Resposta à Radiação , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Humanos , Células MCF-7 , Medicina Tradicional Chinesa/métodos , Doses de Radiação , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/efeitos da radiação , Resultado do Tratamento , Triterpenos/uso terapêutico , Proteína X Associada a bcl-2/metabolismo
18.
Mol Ther Nucleic Acids ; 12: 105-117, 2018 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-30195750

RESUMO

Aberrant insulin-like growth factor I receptor (IGF1R) signaling pathway serves as a well-established target for cancer drug therapy. The intragenic antisense long noncoding RNA (lncRNA) IRAIN, a putative tumor suppressor, is downregulated in breast cancer cells, while IGF1R is overexpressed, leading to an abnormal IGF1R/IRAIN ratio that promotes tumor growth. To precisely target this pathway, we developed an "antisense lncRNA-mediated intragenic cis competition" (ALIC) approach to therapeutically correct the elevated IGF1R/IRAIN bias in breast cancer cells. We used CRISPR-Cas9 gene editing to target the weak promoter of IRAIN antisense lncRNA and showed that in targeted clones, intragenic activation of the antisense lncRNA potently competed in cis with the promoter of the IGF1R sense mRNA. Notably, the normalization of IGF1R/IRAIN transcription inhibited the IGF1R signaling pathway in breast cancer cells, decreasing cell proliferation, tumor sphere formation, migration, and invasion. Using "nuclear RNA reverse transcription-associated trap" sequencing, we uncovered an IRAIN lncRNA-specific interactome containing gene targets involved in cell metastasis, signaling pathways, and cell immortalization. These data suggest that aberrantly upregulated IGF1R in breast cancer cells can be precisely targeted by cis transcription competition, thus providing a useful strategy to target disease genes in the development of novel precision medicine therapies.

19.
Dose Response ; 16(3): 1559325818799561, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30263020

RESUMO

MicroRNAs (miRs), which regulate target gene expression at the post-transcriptional level, play a crucial role in inducing biological effects upon high-dose ionizing radiation. Yet, the miR expression profiles in response to repeated low-dose radiation (LDR) in vivo have not been elucidated. This study investigated the response profiles of 11 miRs with functions involved in metabolism, DNA damage and repair, inflammation, and fibrosis in mouse liver, heart, and testis upon repeated LDR exposure for 4 months. The expression profiles were evaluated using stem-loop quantitative reverse transcription polymerase chain reaction immediately and at 2 months after LDR exposure. The expression profiles varied significantly at both time points. At the organ level, the heart was the most affected, followed by the liver and testis, in which significant miR upregulation related to DNA damage response was found. Metabolism-related miRs decreased in the liver and increased in the testis. The current results showed immediate and long-lasting alterations in the miR expression profiles in response to repeated LDR in different organs.

20.
Mol Med Rep ; 18(3): 3211-3218, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30085342

RESUMO

Chitosan is a linear polysaccharide that is made by treating the chitin shells of shrimp and crustaceans with an alkaline substance, for example sodium hydroxide. Due to its unique physical and chemical properties, chitosan has a wide range of applications in the medical field. Currently, there are no effective treatments for liver fibrosis; therefore, the aim of the present study was to investigate the therapeutic effect of chitosan in a CCl4­induced hepatic fibrosis (HF) rat model. The serum levels of aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP) were measured by ELISA. Collagen (COL) 3 and α­smooth muscle actin (SMA) expression levels in the rat liver were detected by reverse transcription­semiquantitative polymerase chain reaction and western blotting, respectively. The results demonstrated that treatment with chitosan significantly improved HF, by decreasing the serum levels of AST, ALT, and ALP; improving liver histology; and decreasing the expression levels of COL3 and α­SMA. Chitosan may offer an alternative approach for the clinical treatment of HF.


Assuntos
Antioxidantes/uso terapêutico , Quitosana/uso terapêutico , Cirrose Hepática/tratamento farmacológico , Actinas/genética , Actinas/metabolismo , Alanina Transaminase/sangue , Fosfatase Alcalina/sangue , Animais , Antioxidantes/química , Aspartato Aminotransferases/sangue , Tetracloreto de Carbono , Quitosana/química , Colágeno Tipo III/genética , Colágeno Tipo III/metabolismo , Expressão Gênica/efeitos dos fármacos , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/patologia , Masculino , Ratos , Ratos Sprague-Dawley
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