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The chromatin modifier GRAIN WEIGHT 6a (GW6a) enhances rice grain size and yield. However, little is known about its gene network determining grain size. Here, we report that MITOGEN-ACTIVED PROTEIN KINASE 6 (OsMAPK6) and E3 ligase CHANG LI GENG 1 (CLG1) interact with and target GW6a for phosphorylation and ubiquitylation, respectively. Unexpectedly, however, in vitro and in vivo assays reveal that both of the two post-translational modifications stabilize GW6a. Furthermore, we uncover two major GW6a phosphorylation sites (serine142 and threonine186) targeted by OsMAPK6 serving an important role in modulating grain size. In addition, our genetic and molecular results suggest that the OsMAPK6-GW6a and CLG1-GW6a axes are crucial and operate in a non-additive manner to control grain size. Overall, our findings identify a previously unknown mechanism by which phosphorylation and ubiquitylation non-additively stabilize GW6a to enhance grain size, and reveal correlations and interactions of these posttranslational modifications during rice grain development.
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Regulação da Expressão Gênica de Plantas , Oryza , Proteínas de Plantas , Ubiquitinação , Oryza/metabolismo , Oryza/genética , Oryza/crescimento & desenvolvimento , Fosforilação , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/genética , Grão Comestível/metabolismo , Grão Comestível/crescimento & desenvolvimento , Processamento de Proteína Pós-Traducional , Plantas Geneticamente Modificadas , Cromatina/metabolismoRESUMO
[This corrects the article DOI: 10.3892/ol.2019.10694.].
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The present study focused on exploring the inhibitory mechanism of microRNA (miR)-23a in endometrial cancer. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to investigate miR-23a expression in endometrial tissues and endometrial cancer cells. A colony formation assay using crystal violet staining was performed to compare cell proliferation, while wound-healing and Transwell assays were performed to compare cell migration and invasion. Subsequently, bioinformatics and a luciferase reporter gene assay were used to investigate the effect of miR-23a on sine oculis homeobox homolog 1 (SIX1) expression, and the biological function of SIX1 was analyzed. Additionally, a nude mouse tumorigenicity assay was performed to test the inhibitory effect of miR-23a and Taxol® therapy in endometrial cancer. Finally, immunohistochemistry and RT-qPCR were used to explore the association between miR-23a and SIX1 expression in endometrial cancer tissues. miR-23a was underexpressed in endometrial cancer tissues compared with in para-carcinoma tissues, and the overexpression of miR-23a inhibited proliferation and invasion of endometrial cancer cells. Furthermore, SIX1 was demonstrated to be a downstream target of miR-23a, and miR-23a reduced SIX1 expression. Additionally, SIX1 inversely promoted cell proliferation, migration and invasion. In addition, the effects of reduced cell proliferation and increased cell invasion following miR-23a overexpression could be reversed by adding SIX1 to in vitro culture. Furthermore, the inhibitory effect of miR-23a and Taxol therapy, which reduced SIX1 expression in endometrial cancer, was demonstrated in vivo. Finally, a negative association between miR-23a and SIX1 expression was demonstrated in endometrial cancer tissues. The results of the present study revealed that miR-23a may inhibit endometrial cancer development by targeting SIX1.
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BACKGROUND: Cervical cancer is the most common gynecological malignancy with low terminal cure rate, and therefore new therapeutic targets are urgently needed to combat this disease. SMYD2, as an oncogene, is abnormal highly expressed in multiple types of tumors and further affects the occurrence and development, but the potential correlations between SMYD2 expression and cervical cancer progression is still unclear. METHODS: We first used the bioinformatics website to screen the data of cervical cancer in (The Cancer Genome Atlas) TCGA and survival analysis was used to find the different survival rates in the SMYD2 high expression group and low expression group. Through immunohistochemistry, the association between SMYD2 expression and clinical-pathological features of cervical cancer patients was further evaluated. Quantitative PCR and Immunoblot were applied to investigate the relative mRNA and protein expression levels, respectively. In vivo and in vitro experiments were performed to explore the function of SMYD2 in cancer progression. RESULTS: We first found a high expression of SMYD2 in cervical cancer, and survival analysis found that the poorer survival rate in the SMYD2 high expression group than that in the low expression group. Herein, our study demonstrated that the expression of SMYD2 in patients with cervical cancer was associated with FIGO stage, tumor size and further correlated with poor prognosis. Our data further showed that the inhibition of SMYD2 expression in cervical cancer cell line Caski and Siha could dramatically block the proliferation of cervical cancer cells. Additionally, SMYD2-shRNA lentivirus infected remarkably inhibited tumorigenesis in mice compared with the scramble group. CONCLUSIONS: Taken together, this study provides strong evidence of the involvement of SMYD2 in cervical cancer growth and indicates that it could have high potential as a therapeutic target of cervical cancer.
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Cervical cancer is one of the most common malignant neoplasms in gynecology. Protein tyrosine kinase 7 (PTK7) with an inactive kinase domain is an important regulator of multiple Wnt pathways under normal and various pathological conditions and overexpressed in various tumors; however, the clinical and biological significance of PTK7 in cervical cancer is still unknown. In the present study, the protein expression level of PTK7 was detected in clinical cervical cancer patient samples, and the relationship between PTK7 expression and clinicopathological features was analyzed. In addition, the Kaplan-Meier method was performed to estimate the overall survival (OS) and progression-free survival (PFS) of patients to investigate the clinicopathological significance of PTK7 expression. Functional assays demonstrated that knocking down PTK7 might inhibit the ability of cancer cells to proliferate and invade or migrate, both in vivo and in vitro. Thus, PTK7 might serve as a potential target for cervical cancer.
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Biomarcadores Tumorais/genética , Moléculas de Adesão Celular/genética , Receptores Proteína Tirosina Quinases/genética , Neoplasias do Colo do Útero/genética , Animais , Biomarcadores Tumorais/metabolismo , Moléculas de Adesão Celular/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Feminino , Humanos , Camundongos , Receptores Proteína Tirosina Quinases/metabolismo , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologiaRESUMO
BACKGROUND: To evaluate the diagnostic performances of detecting circulating tumor cells (CTCs) and tumor cells in bronchoalveolar lavage fluid (BALF) for peripheral lung cancer. METHODS: A total of 247 patients with lung cancer and 70 cases with benign lung disease were recruited in this study. Peripheral blood and BALF samples were collected, in which the tumor cells were enriched by negative immunomagnetic selection and detected by fluorescence in situ hybridization (FISH) of chromosome enumeration probe 8 (CEP8). The levels of tumor-associated markers (e.g., CEA, CA125, and NSE) in peripheral blood plasma were measured by using electrochemiluminescence. RESULTS: The numbers of CTCs detected in peripheral blood were significantly higher in patients with lung cancer than those with benign lung disease (5.78±0.57 vs. 1.13±0.39, Z=-8.64, P<0.01). Similarly, tumor cells count in BALF of malignancy were higher than that of benign lesions (6.76±0.89 vs. 0.89±0.23, Z=-6.254, P<0.01). However, as for patients with lung cancer and benign lung disease, the numbers of tumor cells in peripheral blood were comparable with those in BALF (both P>0.05). Detecting CTCs and tumor cells in BALF had similar areas under curves (AUC =0.871 and 0.963, respectively; P>0.05) in discriminating benign lesions from lung cancer (sensitivity 83.8% and 92.6%, specificity 86.5% and 99.9%, respectively), both of which were larger than those of NSE, CEA, and CA125 (AUC =0.564, 0.512 and 0.554, respectively; all P<0.05). The diagnostic performances of discriminating benign lesions and lung cancer in BALF and peripheral blood were both in concordance with that of histopathology (kappa values 0.662 and 0.569, respectively; both P<0.001). CONCLUSIONS: Detecting tumor cells in peripheral blood and BALF may sensitive to identify benign and malignant peripheral lung lesions and be of value for early diagnosis of lung cancer.
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Infecções por Coxsackievirus/virologia , Enterovirus Humano A/fisiologia , Infecções por Enterovirus/virologia , Interações Hospedeiro-Patógeno/imunologia , Infecções por Coxsackievirus/imunologia , Infecções por Enterovirus/imunologia , Regulação da Expressão Gênica/imunologia , Regulação Viral da Expressão Gênica , HumanosRESUMO
OBJECTIVE: To study the therapeutic effect of recombinant adeno-associated virus carrying human endostatin gene therapy on endometriosis in mice model. METHODS: Recombinant adeno-associated virus vector carrying human endostatin gene and enhanced green fluorescent proteins gene (rAAV2-endostatin-EGFP) was constructed. Endometrium was from 12 patients with leiomyoma undergoing hysterectomy in Second Hospital, Tianjin Medical University between November and December 2008. Endometriosis models of nude mice were established by transplanting human endometrial fragments intooperitoneal surface. After 1 week, those 60 mice were divided into 3 groups: treatment group including 20 mice injected with rAAV2-endostatin-EGFP to ectopic lesion, control group including 20 mice injected with rAAV2-EGFP to ectopic lesion and blank control group including 20 mice injected with phosphate buffered saline (PBS) to the ectopic lesion. At 1, 2 and 3 weeks after treatment, those mice underwent laparotomy to observe the location and size of ectopic lesion in abdominal cavity. The expression of endostain protein, number of gland, microvessel density (MVD) and vascular endothelial growth factor (VEGF) were measured in ectopic lesions. The serum level of estradiol and progesterone were detected in nude mice among every groups. RESULTS: (1) All endometriosis of nude mice models were established successfully through peritoneum transplanting. After 1 week's treatment, flat lesion nodes, decreased gland number and narrow and atrophy glandular cavity were observed by light microscope. (2) The endostatin gene was transferred into nude mice successfully and expressed effectively. It was observed that endostatin protein expression was shown with enhanced green fluorescent proteins in ectopic lesion. (3) Glands number of ectopic lesion in rAAV2-endostatin-EGFP group (7.8 ± 1.9, 7.0 ± 1.5 and 5.5 ± 1.7) were significantly less than 10.1 ± 1.7, 10.2 ± 2.0 and 9.8 ± 2.4 in rAAV2-EGFP control group and 10.2 ± 2.2, 10.0 ± 2.0 and 9.7 ± 2.2 in PBS control group at 1, 2 and 3 weeks after treatment (all P < 0.05). Glands number of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P < 0.05). (4) MVD of ectopic lesion in rAAV2-endostatin-EGFP group (12.2 ± 1.5, 11.4 ± 2.1 and 9.0 ± 1.4) was significantly less than those at rAAV2-EGFP control group (16.5 ± 1.7, 16.5 ± 1.9 and 16.9 ± 1.9) and PBS control group (16.2 ± 1.6, 16.0 ± 1.6 and 16.3 ± 1.7) at 1, 2 and 3 weeks after treatment (all P < 0.05). MVD of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P < 0.05). (5) The rate and density of VEGF expression at ectopic lesion in rAAV2-endostatin-EGFP group (35%, 30%, 25% and 1.60 ± 0.43, 1.33 ± 0.30, 1.03 ± 0.36) were significantly less than those at rAAV2-EGFP control group (80%, 75%, 85% and 2.43 ± 0.53, 2.43 ± 0.29, 2.66 ± 0.45) and PBS control group (85%, 90%, 90% and 2.36 ± 0.53, 2.64 ± 0.57, 2.53 ± 0.52) at one 1, 2 and 3 weeks after treatment (all P < 0.05). The expression of VEGF at ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P < 0.05). (6) The level of estradial and progesterone in serum of nude mice of rAAV2-endostatin-EGFP group [E(2): (48 ± 7) pmol/L, P: (61 ± 8) nmol/L] did not reach statistical difference when compared with those at rAAV2-EGFP control group [E(2): (50 ± 9) pmol/L, P: (60 ± 10) nmol/L] and PBS control group [E(2): (48 ± 7) pmol/L, P: (58 ± 10) nmol/L, P > 0.05]. CONCLUSIONS: The recombinant adeno-associated virus carrying human endostatin gene therapy could inhibit angiogenesis at endometriotic lesions and not influence steroid level. The antiangiogenic gene therapy might become a novel option for endometriosis.
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Inibidores da Angiogênese/uso terapêutico , Endometriose/terapia , Endostatinas/genética , Terapia Genética/métodos , Inibidores da Angiogênese/genética , Animais , Dependovirus/genética , Modelos Animais de Doenças , Endometriose/genética , Endometriose/metabolismo , Endostatinas/metabolismo , Endostatinas/uso terapêutico , Feminino , Vetores Genéticos , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neovascularização Patológica , Proteínas Recombinantes/uso terapêutico , Recombinação Genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
OBJECTIVE: To assess interleukin (IL)-1alpha, beta and interferon (IFN) gamma expression in macrophages in eutopic and ectopic endometrium of women with endometriosis. METHODS: In situ hybridization was used to examine the expression of IL-1alpha and beta and IFN-gamma in macrophages from eutopic and ectopic endometrium of 40 women with endometriosis and 15 control women. Eutopic endometrial samples were histologically classified into proliferative and secretary phases. Cervical polyp tissue was used as positive control. RESULTS: Expression of IL-1alpha and beta in macrophages from eutopic and ectopic endometrium of women with endometriosis were significantly higher than that in the control group (P < 0.05). Both gene expression in macrophages from ectopic endometrium was higher than that in eutopic endometrium of patients with endometriosis (P < 0.05). The expressions of the two genes were significantly increased in secretory phase of endometrium when compared to that in proliferative phase (P < 0.05). There was no difference in IFN-gamma expression in macrophages of endometium between patients with endometriosis and control (P > 0.05). No cycle dependent variation of the gene expression in the macrophages was found either in endometriosis group or in control group. CONCLUSION: There is a significant increase in the expression of IL-1alpha and beta in macrophages of endometriosis. IL-1 and activated macrophages may play an important role in the development and progression of endometriosis.