RESUMO
BACKGROUND: Emerging evidence has shown that circular RNAs (circRNAs) play a crucial regulatory role in the occurrence and development of cancer. Exploring the roles and mechanisms of circRNAs in tumorigenesis and progression may help to identify new diagnostic markers and therapeutic targets. In the present study, we investigated the role and regulatory mechanism of hsa_circ_0004872 in gastric cancer (GC). METHODS: qRT-PCR was used to determine the expression of hsa_circ_0004872 in GC tissues and cells. EdU, CCK-8, transwell and scratch wound healing assays were used to assess the role of hsa_circ_0004872 in GC cell proliferation, invasion and migration, respectively. Subcutaneous and tail vein tumor injections in nude mice were used to assess the role of hsa_circ_0004872 in vivo. RIP assay, biotin-coupled probe pull-down assay, FISH and luciferase reporter assay were performed to confirm the relationship between hsa_circ_0004872 and the identified miRNA. ChIP assay, luciferase reporter assay and western blot were used to determine the direct binding of Smad4 to the promoter of the ADAR1 gene. RESULTS: In this study, we found that hsa_circ_0004872 was dramatically downregulated in GC tissues compared with adjacent noncancerous tissues. The expression level of hsa_circ_0004872 was associated with tumor size and local lymph node metastasis. Enforced expression of hsa_circ_0004872 inhibited the proliferation, invasion and migration of GC cells, whereas knockdown of hsa_circ_0004872 had the opposite effects. Nude mice experiments showed that ectopic expression of hsa_circ_0004872 dramatically inhibited tumor growth and metastasis in vivo. Moreover, we demonstrated that hsa_circ_0004872 acted as a "molecular sponge" for miR-224 to upregulate the expression of the miR-224 downstream targets p21 and Smad4. Importantly, we found that the RNA-editing enzyme ADAR1 inhibited hsa_circ_0004872 expression and further led to the upregulation of miR-224. Smad4, the downstream target of miR-224, could further affect hsa_circ_0004872 levels by directly binding to the promoter region of ADAR1 to inhibit ADAR1 expression. CONCLUSIONS: Our findings showed that hsa_circ_0004872 acted as a tumor suppressor in GC by forming a negative regulatory loop consisting of hsa_circ_0004872/miR-224/Smad4/ADAR1. Thus, hsa_circ_0004872 may serve as a potential biomarker and therapeutic target for GC.
Assuntos
Adenosina Desaminase/metabolismo , Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , RNA Circular/genética , Proteínas de Ligação a RNA/metabolismo , Proteína Smad4/metabolismo , Neoplasias Gástricas/patologia , Adenosina Desaminase/genética , Animais , Apoptose , Biomarcadores Tumorais/genética , Movimento Celular , Proliferação de Células , Progressão da Doença , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Prognóstico , Proteínas de Ligação a RNA/genética , Proteína Smad4/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Taxa de Sobrevida , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND The aims of this preliminary study were to evaluate contrast-enhanced ultrasound (CEUS) imaging and the therapeutic response of enlarged superficial lymph nodes in patients with lymphoma before and after chemotherapy and to determine the most useful CEUS response parameters. MATERIAL AND METHODS Forty-three patients with lymphoma, with 43 enlarged superficial lymph nodes, underwent CEUS and conventional ultrasound (US), before treatment and after the first three cycles of chemotherapy. Clinical responses included overall response (OR) and no response (NR). Imaging parameters by time-intensity curve (TIC) included basic intensity (B), wash-out slope and/or decent slope (K), wash-in slope or rise slope (C), time to peak (TTP), area under the gamma curve (Area), arrive time(ATM), peak intensity (PI), change of peak intensity (I) were compared. And receiver operating characteristic (ROC) curve analysis was operated. RESULTS Quantitative parameters of CEUS before and after the first three cycles of chemotherapy showed a significant difference in the AreaΔ, PID, and IΔ in the OR group compared with NR group (P<0.05). There was a significant difference in the Cpre, Areain, PIin, Iin, AreaΔ, PIΔ, and IΔ in the OR group compared with NR group (P<0.05). The effectiveness of the therapeutic response was predicted by the CEUS parameters of IΔ (P<0.05). And ΔArea has the highest diagnostic performance of ineffectiveness. CONCLUSIONS The findings of this study have shown that quantitative analysis by CEUS may be a useful, and objective, imaging method for the evaluation of the therapeutic response of enlarged superficial lymph nodes in lymphoma before and after chemotherapy.
Assuntos
Linfonodos/diagnóstico por imagem , Linfoma/diagnóstico por imagem , Ultrassonografia/métodos , Adulto , Idoso , Meios de Contraste , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dados Preliminares , Curva ROCRESUMO
N6-methyladenosine (m6A) is the predominant post-transcriptional RNA modification in eukaryotes and plays a pivotal regulatory role in various aspects of RNA fate determination, such as mRNA stability, alternative splicing, and translation. Dysregulation of the critical m6A methyltransferase METTL3 is implicated in tumorigenesis and development. Here, this work showed that METTL3 is upregulated in gastric cancer tissues and is associated with poor prognosis. METTL3 methylates the A2318 site within the coding sequence (CDS) region of STAT5A. IGF2BP2 recognizes and binds METTL3-mediated m6A modification of STAT5A through its GXXG motif in the KH3 and KH4 domains, leading to increased stability of STAT5A mRNA. In addition, both METTL3 and IGF2BP2 are positively correlated with STAT5A in human gastric cancer tissue samples. Helicobacter pylori infection increased the expression level of METTL3 in gastric cancer cells, thereby leading to the upregulation of STAT5A. Functional studies indicated that STAT5A overexpression markedly enhances the proliferation and migration of GC cells, whereas STAT5A knockdown has inhibitory effects. Further nude mouse experiments showed that STAT5A knockdown effectively inhibits the growth and metastasis of gastric cancer in vivo. Moreover, as a transcription factor, STAT5A represses KLF4 transcription by binding to its promoter region. The overexpression of KLF4 can counteract the oncogenic impact of STAT5A. Overall, this study highlights the crucial role of m6A in gastric cancer and provides potential therapeutic targets for gastric cancer.
Assuntos
Adenosina , Proliferação de Células , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like , Metiltransferases , Camundongos Nus , Proteínas de Ligação a RNA , Fator de Transcrição STAT5 , Neoplasias Gástricas , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologia , Neoplasias Gástricas/metabolismo , Humanos , Adenosina/análogos & derivados , Adenosina/metabolismo , Metiltransferases/metabolismo , Metiltransferases/genética , Fator de Transcrição STAT5/metabolismo , Fator de Transcrição STAT5/genética , Animais , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Camundongos , Proteínas de Ligação a RNA/metabolismo , Proteínas de Ligação a RNA/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Masculino , Feminino , Infecções por Helicobacter/genética , Infecções por Helicobacter/metabolismo , Infecções por Helicobacter/patologia , Movimento Celular/genética , Camundongos Endogâmicos BALB C , Helicobacter pylori/genética , Proteínas Supressoras de TumorRESUMO
Deubiquitinase (DUB) dysregulation is closely associated with multiple diseases, including tumors. In this study, we used data from The Cancer Genome Atlas and Gene Expression Omnibus databases to analyze the expression of 51 ubiquitin-specific proteases (USPs) in gastric cancer (GC) tissues and adjacent non-neoplastic tissues. The Kaplan-Meier Plotter database was used to analyze the association of the differentially expressed USPs with the overall survival of patients with GC. The results showed that five USPs (USP5, USP10, USP13, USP21, and USP35) were highly expressed in GC tissues and were associated with poor prognosis in patients with GC. Because the epithelial-mesenchymal transition enables epithelial cells to acquire mesenchymal features and contributes to poor prognosis, we investigated whether these USPs had regulatory effects on the key epithelial-mesenchymal transition transcription factor Snail1. Our results showed that USP35 exhibited the most significant regulation on Snail1. Overexpression of USP35 increased and its knockdown decreased Snail1 protein levels. Mechanistically, USP35 interacted with Snail1 and removed its polyubiquitinated chain, thereby increasing its stability. Furthermore, USP35 promoted the invasion and migration of GC cells depending on its DUB activity. USP35 knockdown exhibited the opposite effect. Snail1 depletion partially abrogated the biological effects of USP35. Experiments using nude mouse tail vein injections indicated that wild-type USP35, but not the catalytically inactive USP35-C450A mutant, dramatically enhanced cell colonization and tumorigenesis in the lungs of mice. In addition, USP35 positively correlated with Snail1 expression in clinical GC tissues. Helicobacter pylori infection increased USP35 and Snail1 expression levels. Altogether, we found that USP35 can deubiquitinate Snail1 and increase its expression, thereby contributing to the malignant progression of GC. Therefore, USP35 may serve as a viable target for GC treatment.
Assuntos
Endopeptidases , Infecções por Helicobacter , Fatores de Transcrição da Família Snail , Neoplasias Gástricas , Animais , Humanos , Camundongos , Carcinogênese , Transformação Celular Neoplásica , Endopeptidases/genética , Camundongos Nus , Neoplasias Gástricas/genética , Ubiquitina Tiolesterase/genética , Proteases Específicas de Ubiquitina/genética , Fatores de Transcrição da Família Snail/genéticaRESUMO
The reversible post-translational modifications of protein ubiquitination and deubiquitination play a crucial regulatory role in cellular homeostasis. Deubiquitinases (DUBs) are responsible for the removal of ubiquitin from the protein substrates. The dysregulation of the DUBs may give rise to the occurrence and development of tumors. In this study, we investigated the gastric cancer (GC) data from the TCGA and GEO databases and found that ubiquitin-specific protease USP13 was significantly up-regulated in GC samples. The higher expression of USP13 was associated with the worse prognosis and shorter overall survival (OS) of GC patients. Enforced expression of USP13 in GC cells promoted the cell cycle progression and cell proliferation in an enzymatically dependent manner. Conversely, suppression of USP13 led to GC cell cycle arrest in G1 phase and the inhibition of cell proliferation. Nude mouse experiments indicated that depletion of USP13 in GC cells dramatically suppressed tumor growth in vivo. Mechanistically, USP13 physically bound to the N-terminal domain of cyclin D1 and removed its K48- but not K63-linked polyubiquitination chain, thereby stabilizing and increasing cyclin D1. Furthermore, re-expression of cyclin D1 partially reversed the cell cycle arrest and cell proliferation inhibition induced by USP13 depletion in GC cells. Additionally, USP13 protein abundance was positively correlated with the protein level of cyclin D1 in human GC tissues. Taken together, our data demonstrate that USP13 deubiquitinates and stabilizes cyclin D1, thereby promoting cell cycle progression and cell proliferation in GC. These findings suggest that USP13 might be a promising therapeutic target for the treatment of GC.
Assuntos
Neoplasias Gástricas , Animais , Camundongos , Humanos , Neoplasias Gástricas/patologia , Ciclina D1/genética , Ciclina D1/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Fase G1 , Proteases Específicas de Ubiquitina/genética , Ciclo CelularRESUMO
Purpose: To evaluate and compare the predictive performance of different deep learning models using gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced MRI in predicting microvascular invasion (MVI) in hepatocellular carcinoma. Methods: The data of 233 patients with pathologically confirmed hepatocellular carcinoma (HCC) treated at our hospital from June 2016 to June 2021 were retrospectively analyzed. Three deep learning models were constructed based on three different delineate methods of the region of interest (ROI) using the Darwin Scientific Research Platform (Beijing Yizhun Intelligent Technology Co., Ltd., China). Manual segmentation of ROI was performed on the T1-weighted axial Hepatobiliary phase images. According to the ratio of 7:3, the samples were divided into a training set (N=163) and a validation set (N=70). The receiver operating characteristic (ROC) curve was used to evaluate the predictive performance of three models, and their sensitivity, specificity and accuracy were assessed. Results: Among 233 HCC patients, 109 were pathologically MVI positive, including 91 men and 18 women, with an average age of 58.20 ± 10.17 years; 124 patients were MVI negative, including 93 men and 31 women, with an average age of 58.26 ± 10.20 years. Among three deep learning models, 2D-expansion-DL model and 3D-DL model showed relatively good performance, the AUC value were 0.70 (P=0.003) (95% CI 0.57-0.82) and 0.72 (P<0.001) (95% CI 0.60-0.84), respectively. In the 2D-expansion-DL model, the accuracy, sensitivity and specificity were 0.7143, 0.739 and 0.688. In the 3D-DL model, the accuracy, sensitivity and specificity were 0.6714, 0.800 and 0.575, respectively. Compared with the 3D-DL model (based on 3D-ResNet), the 2D-DL model is smaller in scale and runs faster. The frames per second (FPS) for the 2D-DL model is 244.7566, which is much larger than that of the 3D-DL model (73.3374). Conclusion: The deep learning model based on Gd-EOB-DTPA-enhanced MRI could preoperatively evaluate MVI in HCC. Considering that the predictive performance of 2D-expansion-DL model was almost the same as the 3D-DL model and the former was relatively easy to implement, we prefer the 2D-expansion-DL model in practical research.
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Occult breast cancer is an uncommon type of breast cancer. Metastases of occult breast cancer to other tissues are rather rare. We present a rare case of thyroid metastases in a 46-year-old woman who underwent occult breast cancer. The first ultrasound (US) examination of the thyroid showed that the left lobe was enlarged but had normal thyroid function. At first, this case was misdiagnosed as thyroiditis based on the thyroid US features. However, the cytological and histological results showed that nests of the neoplastic cells were found. Further immunohistochemistry results confirmed that these neoplasms were derived from breast tissue. Analysis using the successive US scans revealed that the sizes and echo of the thyroid repeatedly changed after the radiotherapy and chemotherapy treatment. To our knowledge, this is the first reported case of occult breast carcinoma presenting with thyroid metastases. This case can easily be misdiagnosed as thyroiditis due to the metastasis area not manifesting as regular suspicious nodules or diffused punctate calcifications.
RESUMO
Gastric cancer (GC) is one of the most common malignancies in the world and ranks third in terms of cancer-related deaths. The catalytically inactive pseudophosphatase STYX (serine/threonine/tyrosine interacting protein) is a member of the protein tyrosine phosphatase family. It has been recently reported that STYX functions as a potential oncogene in different types of cancers. However, the potential role and regulatory mechanism of STYX in GC remains unknown. In this study, we find that STYX is highly expressed in GC tissues compared with adjacent noncancerous tissues and closely correlates with the prognosis of GC patients. STYX overexpression facilitates the proliferation and migration in GC cells, whereas STYX knockdown has the opposite effects. Nude mice experiments indicate that STYX knockdown in GC cells dramatically suppresses the tumor growth and lung metastasis in vivo. Mechanically, our results suggest that STYX interacts with the F-box protein FBXO31 and disrupts the degradation function of FBXO31 to its target proteins CyclinD1 and Snail1, thereby increasing the level of CyclinD1 and Snail1 in GC. STYX-mediated biological changes can be reversed by the co-expression of STYX and FBXO31 in GC cells. In addition, transcription factor c-Jun can enhance the expression of STYX in GC. The expression of STYX can also be induced by Helicobacter pylori (H. pylori) infection in c-Jun-dependent manner. Together, our present study suggests that STYX plays an oncogenic role in GC by inhibiting FBXO31 function and represents a potential therapeutic target and prognostic biomarker in GC.
Assuntos
Proteínas F-Box , Helicobacter pylori , Neoplasias Gástricas , Animais , Linhagem Celular Tumoral , Proliferação de Células , Proteínas F-Box/genética , Proteínas F-Box/metabolismo , Regulação Neoplásica da Expressão Gênica , Helicobacter pylori/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Camundongos , Camundongos Nus , Neoplasias Gástricas/patologia , Proteínas Supressoras de Tumor/metabolismoRESUMO
Background: To evaluate the safety and efficacy of radiofrequency ablation (RFA), microwave ablation (MWA), and laser ablation (LA) for the treatment of cervical metastatic lymph nodes (CMLNs) of papillary thyroid carcinoma (PTC). Methods: The Pubmed, EMBASE, Web of Science, and Cochrane Library databases were searched for studies on the safety and efficacy of thermal ablations (RFA, MWA, and LA) for the treatment of CMLNs of PTC until March 30, 2022. A review of 334 potential papers identified 17 eligible papers including 312 patients. Fixed-effects model or random-effects model was used to evaluate the pooled proportions of volume reduction rate (VRR), complete disappearance, and recurrence, and pooled estimates of changes in the largest diameter, volume, and serum Tg after ablation. The pooled proportions of overall and major complications were calculated. Subgroup analysis based on treatment modalities. The heterogeneity among studies was analyzed by using Q statistics and inconsistency index I2 . MINORS scale was used to evaluate the quality of the studies. Results: 17 eligible studies were finally identified, including 312 patients and 559 CMLNs. The pooled proportions of VRR, complete disappearance and recurrence of CMLNs were 91.28% [95% confidence interval (CI): 86.60-95.97%], 67.9% [95% CI: 53.1-81.1%] and 7.8% [95%CI: 3.0-14.1%], respectively. The pooled estimates of changes in the largest diameter, volume and serum Tg were 8.12 mm [95%CI: 6.78-9.46 mm], 338.75 mm3 [95%CI: 206.85 -470.65 mm3] and 5.96 ng/ml [95%CI: 3.68-8.24 ng/ml], respectively. The pooled proportions of overall and major complications were 2.9% [95%CI: 0.3-7.1%] and 0.3% [95%CI: 0-1.9%], respectively. Significant between-study heterogeneity was observed for complete disappearance (P<0.01, I2 =88.6%), VRR (P<0.001, I2 =99.9%), recurrence (P=0.02, I2 =47.76%), overall complications (P<0.02, I2 =44.8%), and changes in the largest diameter (P < 0.001, I2 =82.6%), volume (P<0.001, I2 =97.0%), and serum Tg (P < 0.001, I2 =93.7%). Subgroup analysis showed heterogeneity of the VRR among the treatment modality (I2 range: 84.4-100%). The VRR of MWA was the highest (97.97%), followed by RFA (95.57%) and LA (84.46%) (P < 0.001). Conclusion: All thermal ablations were safe and effective for the treatment of CMLNs of PTC. However, each treatment had significant heterogeneity in VRR. Compared with RFA and MWA, LA was less effective in reducing the volume of CMLNs of PTC.
Assuntos
Ablação por Cateter , Ablação por Radiofrequência , Neoplasias da Glândula Tireoide , Ablação por Cateter/efeitos adversos , Humanos , Linfonodos/cirurgia , Ablação por Radiofrequência/efeitos adversos , Câncer Papilífero da Tireoide/cirurgia , Neoplasias da Glândula Tireoide/etiologia , Neoplasias da Glândula Tireoide/cirurgiaRESUMO
BACKGROUND: Chronic kidney disease (CKD) has become a major public health issue. Meanwhile, renal fibrosis caused by diabetic nephropathy can lead to CKD, regardless of the initial injury. It has been previously reported that silibinin or valsartan could relieve the severity of renal fibrosis. However, the effect of silibinin in combination with valsartan on renal fibrosis remains unclear. MATERIAL AND METHODS: Proximal tubular cells (HK-2) were treated with TGF-ß1 (5 ng/mL) to mimic in vitro model of fibrosis. The proliferation of HK-2 cells was tested by CCK-8. Epithelial-mesenchymal transition (EMT) and inflammation-related gene and protein expressions in HK-2 cells were measured by qRT-PCR and Western-blot, respectively. ELISA was used to test the level of TNF-αNF-A. Additionally, HFD-induced renal fibrosis mice model was established to investigate the effect of silibinin in combination with valsartan on renal fibrosis in vivo. RESULTS: Silibinin significantly increased the anti-fibrosis effect of valsartan in TGF-ß1-treated HK-2 cells via inhibition of TGF-ß1 signaling pathway. Furthermore, silibinin significantly enhanced the anti-fibrosis effect of valsartan on HFD-induced renal fibrosis in vivo through inactivation of TGF-ß1 signaling pathway. CONCLUSION: These data indicated that silibinin markedly increased anti-fibrosis effect of valsartan in vitro and in vivo. Thus, silibinin in combination with valsartan may act as a potential novel strategy to treat renal fibrosis caused by diabetic nephropathy.
Assuntos
Nefropatias Diabéticas/tratamento farmacológico , Túbulos Renais Proximais/efeitos dos fármacos , Silibina/farmacologia , Fator de Crescimento Transformador beta1/metabolismo , Valsartana/farmacologia , Animais , Linhagem Celular , Nefropatias Diabéticas/complicações , Modelos Animais de Doenças , Sinergismo Farmacológico , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Fibrose/prevenção & controle , Humanos , Túbulos Renais Proximais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Insuficiência Renal Crônica/prevenção & controle , Transdução de Sinais/efeitos dos fármacos , Silibina/administração & dosagem , Valsartana/administração & dosagemRESUMO
OBJECTIVE: Application of rapid on-site evaluation (ROSE) for thyroid fine needle aspiration (FNA) is controversial. Therefore, ROSE has not been universally applied. This study aimed to evaluate the value of ROSE for ultrasound-guided thyroid FNA. METHODS: A total of 997 patients with 1103 suspicious thyroid nodules had ultrasound-guided FNA performed from January 2016 to February 2018. There were 513 nodules with ROSE and 590 nodules without ROSE. The cytological nondiagnostic rate, needle passes, and procedural times of thyroid FNA with or without ROSE were compared. The nondiagnostic rates of subsets of suspicious thyroid nodules were further compared. RESULTS: There was no significant effect of ROSE on the nondiagnostic rate of FNA. However, FNA with ROSE significantly reduced the numbers of sub-centimeter, mixed solid-cystic, macrocalcified, and hypervascular nodules. There was a significantly smaller number of needle passes and less procedural times with ROSE than without ROSE. There was no significant difference in the complication rate of FNA with and without ROSE. CONCLUSION: ROSE for thyroid FNA reduces the number of needle passes and procedural times. ROSE has a higher clinical application value in subsets of thyroid nodules, which tend to be difficult to diagnose with FNA.