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1.
FEBS Open Bio ; 14(7): 1205-1217, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38872260

ABSTRACT

Clear cell renal cell carcinoma (ccRCC) accounts for approximately 75-80% of all patients with renal cell carcinoma. Despite its prevalence, little is known regarding the key components involved in ccRCC metastasis. In this study, scRNA-seq analysis was employed to classify CD8+ T cells into four sub-clusters based on their genetic profiles and immunofluorescence experiments were used to validate two key clusters. Through gene set enrichment analysis, these newly identified sub-clusters were found to exhibit distinct biological characteristics. Notably, TYMP, TOP2A, CHI3L2, CDKN3, CENPM, and RZH2 were highly expressed in these sub-clusters, indicating a correlation with poor prognosis. Among these sub-clusters, CD8+ T cells (MT-ND4) were identified as potentially playing a critical role in mediating ccRCC metastasis. These results contribute to our understanding of CD8+ T cell heterogeneity in ccRCC and shed light on the mechanisms underlying the loss of immune response against cancer.


Subject(s)
CD8-Positive T-Lymphocytes , Carcinoma, Renal Cell , Kidney Neoplasms , Carcinoma, Renal Cell/pathology , Carcinoma, Renal Cell/immunology , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/metabolism , Humans , Kidney Neoplasms/pathology , Kidney Neoplasms/immunology , Neoplasm Metastasis , Prognosis , Gene Expression Regulation, Neoplastic
2.
Front Genet ; 14: 1222064, 2023.
Article in English | MEDLINE | ID: mdl-37396044
3.
Mediators Inflamm ; 2023: 9810733, 2023.
Article in English | MEDLINE | ID: mdl-37273451

ABSTRACT

Ferroptosis resistance is vital for B cell development, especially in inflammatory diseases, yet the underlying mechanism is still unclear. In this study, based on the scRNA-seq technique and flow cytometry, we discovered a proportion of neutrophils exhibited upregulated expression of the IL-6 and correlated with the expression of IL-6 receptor and SLC7A11 from B cells in lupus kidney. Moreover, we identified that in lupus kidney, neutrophils could provide IL-6 to facilitate ferroptosis resistance in B cells via SLC7A11, and inhibition of SLC7A11 could significantly enhance ferroptosis in B cells and could decrease B cell proliferation. This study helps understand the crosstalk between neutrophils and B cells in the kidney in the development of lupus.


Subject(s)
Ferroptosis , Interleukin-6 , Lupus Nephritis , Humans , Kidney , Neutrophils , B-Lymphocytes
4.
J Magn Reson Imaging ; 54(2): 421-428, 2021 08.
Article in English | MEDLINE | ID: mdl-33590944

ABSTRACT

BACKGROUND: Myocardial injury has been found using magnetic resonance imaging in recovered coronavirus disease 2019 (COVID-19) patients unselected or with ongoing cardiac symptoms. PURPOSE: To evaluate for the presence of myocardial involvement in recovered COVID-19 patients without cardiovascular symptoms and abnormal serologic markers during hospitalization. STUDY TYPE: Prospective. POPULATION: Twenty-one recovered COVID-19 patients and 20 healthy controls (HC). FIELD STRENGTH/SEQUENCE: 3.0 T, cine, T2-weighted imaging, T1 mapping, and T2 mapping. ASSESSMENT: Cardiac ventricular function includes end-diastolic volume, end-systolic volume, stroke volume, cardiac output, left ventricle (LV) mass, and ejection fraction (EF) of LV and right ventricle (RV), and segmental myocardial T1 and T2 values were measured. STATISTICAL TESTS: Student's t-test, univariate general linear model test, and chi-square test were used for analyses between two groups. Ordinary one-way analyses of variance or Kruskal-Wallis H test were used for analyses between three groups, followed by post-hoc analyses. RESULTS: Fifteen (71.43%) COVID-19 patients had abnormal magnetic resonance findings, including raised myocardial native T1 (5, 23.81%) and T2 values (10, 47.62%), decreased LVEF (1, 4.76%), and RVEF (2, 9.52%). The segmental myocardial T2 value of COVID-19 patients (49.20 [46.1, 54.6] msec) was significantly higher than HC (48.3 [45.2, 51.7] msec) (P < 0.001), while the myocardial native T1 value showed no significant difference between COVID-19 patients and HC. The myocardial T2 value of serious COVID-19 patients (52.5 [48.1, 57.1] msec) was significantly higher than unserious COVID-19 patients (48.8 [45.9, 53.8] msec) and HC (48.3 [45.2, 51.7]) (P < 0.001). COVID-19 patients with abnormally elevated D-dimer, C-reactive protein, or lymphopenia showed higher myocardial T2 values than without (all P < 0.05). DATA CONCLUSION: Cardiac involvement was observed in recovered COVID-19 patients with no preexisting cardiovascular disease, no cardiovascular symptoms, and elevated serologic markers of myocardial injury during the whole course of COVID-19. LEVEL OF EVIDENCE: 1 TECHNICAL EFFICACY: Stage 5.


Subject(s)
COVID-19 , Heart , Humans , Magnetic Resonance Imaging, Cine , Myocardium , Predictive Value of Tests , Prospective Studies , SARS-CoV-2 , Stroke Volume , Ventricular Function, Left
5.
J Cancer ; 11(15): 4397-4405, 2020.
Article in English | MEDLINE | ID: mdl-32489458

ABSTRACT

Although the roles and underlying mechanisms of other PDK family members (i.e., PDK1, PDK2 and PDK3) in tumor progression have been extensively investigated and are well understood, the functions and underlying molecular mechanisms of pyruvate dehydrogenase kinase 4 (PDK4) in the tumorigenesis and progression of various cancers [including hepatocellular carcinoma (HCC)] remain largely unknown. In this study, we examined the expression profile of PDK4 in HCC clinical tissue specimens and the roles of PDK4 in the proliferation, tumorigenicity, motility and invasion of HCC cells. The immunohistochemistry (IHC) and quantitative real-time PCR (qRT-PCR) results revealed that PDK4 was significantly downregulated in the cohort of HCC clinical specimens. Additionally, PDK4 protein was found in both the nucleus and cytoplasm of HCC cells based on an immunofluorescence (ICC) assay, and PDK4 protein was also found in the nucleus and cytoplasm of cancer cells contained in HCC clinical specimens based on IHC. The CCK-8 assay and cell colony formation assay demonstrated that stable depletion of endogenous PDK4 by lentivirus-mediated RNA interference (RNAi) markedly promoted the proliferation of HCC cell lines (i.e., BEL-7402 and BEL-7404 cells) in vitro, while PDK4 silencing significantly enhanced the tumorigenic ability of BEL-7404 cells in vivo. In addition to enhance proliferation and tumorigenesis induced by PDK4 silencing, additional studies demonstrated that knockdown of PDK4 led to increase migration and invasion of BEL-7402 and BEL-7404 cells in vitro. Taken together, these findings suggest that the loss of PDK4 expression contributes to HCC malignant progression.

6.
Anal Chim Acta ; 1034: 144-152, 2018 Nov 30.
Article in English | MEDLINE | ID: mdl-30193628

ABSTRACT

In our study, the carbon nanodots (CDs) were synthesized by one-step solvothermal method using resorcinol as the only presusor. The obtained CDs contained abundant unsaturated oxygen-containing groups resulting from the surface oxidation. A novel, simple, and real-time fluorescent assay for the detection of water in various organic solvents was thus established by reducing the surface oxidation states. Excellent reversibility can be readily achieved by the external stimulus water and N,N'-dicyclohexylcarbodiimide (DCC). The water-induced sensitive (limit of detection = 0.006%, v/v, in ethanol) and ultrafast (<1 s) response in emission properties was capable of water determination in spirit samples in both solution and solid-state paper test strips.

7.
Sci Rep ; 6: 34501, 2016 Sep 30.
Article in English | MEDLINE | ID: mdl-27687577

ABSTRACT

Targeted disruption of Cripto-1 in mice caused embryonic lethality at E7.5, whereas we unexpectedly found that ectopic Cripto-1 expression in mouse embryos also led to embryonic lethality, which prompted us to characterize the causes and mechanisms underlying embryonic death due to ectopic Cripto-1 expression. RCLG/EIIa-Cre embryos displayed complex phenotypes between embryonic day 14.5 (E14.5) and E17.5, including fatal hemorrhages (E14.5-E15.5), embryo resorption (E14.5-E17.5), pale body surface (E14.5-E16.5) and no abnormal appearance (E14.5-E16.5). Macroscopic and histological examination revealed that ectopic expression of Cripto-1 transgene in RCLG/EIIa-Cre embryos resulted in lethal cardiac defects, as evidenced by cardiac malformations, myocardial thinning, failed assembly of striated myofibrils and lack of heartbeat. In addition, Cripto-1 transgene activation beginning after E8.5 also caused the aforementioned lethal cardiac defects in mouse embryos. Furthermore, ectopic Cripto-1 expression in embryonic hearts reduced the expression of cardiac transcription factors, which is at least partially responsible for the aforementioned lethal cardiac defects. Our results suggest that hemorrhages and cardiac abnormalities are two important lethal factors in Cripto-1 transgenic mice. Taken together, these findings are the first to demonstrate that sustained Cripto-1 transgene expression after E11.5 causes fatal hemorrhages and lethal cardiac defects, leading to embryonic death at E14.5-17.5.

8.
PLoS Genet ; 12(10): e1006308, 2016 Oct.
Article in English | MEDLINE | ID: mdl-27711113

ABSTRACT

miR-155 plays critical roles in numerous physiological and pathological processes, however, its function in the regulation of blood glucose homeostasis and insulin sensitivity and underlying mechanisms remain unknown. Here, we reveal that miR-155 levels are downregulated in serum from type 2 diabetes (T2D) patients, suggesting that miR-155 might be involved in blood glucose control and diabetes. Gain-of-function and loss-of-function studies in mice demonstrate that miR-155 has no effects on the pancreatic ß-cell proliferation and function. Global transgenic overexpression of miR-155 in mice leads to hypoglycaemia, improved glucose tolerance and insulin sensitivity. Conversely, miR-155 deficiency in mice causes hyperglycemia, impaired glucose tolerance and insulin resistance. In addition, consistent with a positive regulatory role of miR-155 in glucose metabolism, miR-155 positively modulates glucose uptake in all cell types examined, while mice overexpressing miR-155 transgene show enhanced glycolysis, and insulin-stimulated AKT and IRS-1 phosphorylation in liver, adipose tissue or skeletal muscle. Furthermore, we reveal these aforementioned phenomena occur, at least partially, through miR-155-mediated repression of important negative regulators (i.e. C/EBPß, HDAC4 and SOCS1) of insulin signaling. Taken together, these findings demonstrate, for the first time, that miR-155 is a positive regulator of insulin sensitivity with potential applications for diabetes treatment.


Subject(s)
Diabetes Mellitus, Type 2/genetics , Hyperglycemia/genetics , Insulin Resistance/genetics , Insulin/genetics , MicroRNAs/genetics , Adipose Tissue/metabolism , Adipose Tissue/pathology , Animals , Cell Proliferation/genetics , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/metabolism , Gene Expression Regulation , Glucose/metabolism , Humans , Hyperglycemia/blood , Hyperglycemia/pathology , Insulin/metabolism , Insulin Receptor Substrate Proteins/genetics , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Mice , Mice, Transgenic , MicroRNAs/biosynthesis , Muscle, Skeletal/metabolism , Muscle, Skeletal/pathology , Proto-Oncogene Proteins c-akt/genetics
9.
Cancer Med ; 5(12): 3489-3499, 2016 12.
Article in English | MEDLINE | ID: mdl-27774777

ABSTRACT

MicroRNAs (miRNAs) may act as either tumor suppressors or oncogenes in various types of cancers. Previous studies have indicated that miR-17-5p is involved in the initiation and development of human tumors. However, its mechanism and function in nasopharyngeal carcinoma (NPC) remain largely unclear. In this study, we evaluated the expression profiles of miR-17-5p and p21 in NPC cell lines and tissues by quantitative real-time PCR (qRT-PCR). For the analysis, we have established a stable overexpression or depletion of miR-17-5p NPC cell lines for analyzing the effects of cell proliferation by MTT, colony formation, and cell cycle assay. A nude mice xenograft model was used to verify the tumor growth in vivo. MiR-17-5p was overexpressed, whereas the expression of p21 was downregulated in NPC cell lines and tissues. The miR-17-5p expression level was inversely correlated with the p21 mRNA level in NPC samples. Furthermore, analysis of 2-ΔΔCt value in 81 NPC patients suggested that the elevated expression level of miR-17-5p or the downregulated expression level of p21 was significantly correlated with tumor size (T classification) and tumor stage, and Kaplan-Meier survival analysis revealed a correlation between miR-17-5p or p21 expression level and overall survival times in 81 NPC patients. MiR-17-5p promoted cell growth in vivo and in vitro by directly targeting p21. Our results indicate that miR-17-5p can promote the occurrence of NPC and it may serve as a potential novel diagnostic maker or therapeutic target for NPC in the future.


Subject(s)
Carcinoma/genetics , Cell Transformation, Neoplastic/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , Nasopharyngeal Neoplasms/genetics , RNA Interference , p21-Activated Kinases/genetics , 3' Untranslated Regions , Animals , Carcinoma/metabolism , Carcinoma/pathology , Cell Line, Tumor , Cell Proliferation , Disease Models, Animal , Humans , Mice , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/metabolism , Nasopharyngeal Neoplasms/pathology , Retinoblastoma Protein/metabolism , Signal Transduction , Xenograft Model Antitumor Assays
10.
Oncoimmunology ; 5(3): e1086060, 2016 Mar.
Article in English | MEDLINE | ID: mdl-27141341

ABSTRACT

There is an urgent need for more potent and safer approaches to eradicate cancer stem cells (CSCs) for curing cancer. In this study, we investigate cancer-killing activity (CKA) of cytokine-induced killer (CIK) cells against CSCs of hepatocellular carcinoma (HCC). To visualize CSCs in vitro by fluorescence imaging, and image and quantify CSCs in tumor xenograft-bearing mice by bioluminescence imaging, HCC cells were engineered with CSC detector vector encoding GFP and luciferase controlled by Nanog promoter. We found that CIK cells have a strong CKA in vitro against putative CSCs of HCC, as shown by tumorsphere formation and time-lapse imaging. Additionally, time-lapse recording firstly revealed that putative CSCs were attacked simultaneously by many CIK cells and finally eradicated by CIK cells, indicating the necessity of achieving sufficient effector-to-target ratios. We firstly illustrated that anti-NKG2D antibody blocking partially but significantly inhibited CKA of CIK cells against putative CSCs. More importantly, intravenous infusion of CIK cells remarkably delayed tumor growth in mice with a significant decrease in putative CSC number monitored by bioluminescence imaging. Taken together, these findings demonstrate CKA of CIK cells against putative CSCs of HCC, at least in part, by NKG2D-ligands recognition.

11.
Medicine (Baltimore) ; 95(7): e2713, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26886608

ABSTRACT

The miR-17-92 cluster and its 6 different mature microRNAs, including miR-17, miR-18a, miR-19a, miR-20a, miR-19b-1, and miR-92a, play important roles in embryo development, immune system, kidney and heart development, adipose differentiation, aging, and tumorigenicity. Currently, increasing evidence indicates that some members of miR-17-92 cluster may be critical players in spermatogenesis, including miR-17, miR-18a, and miR-20a. However, the roles and underlying mechanisms of miR-17-92 in spermatogenesis remain largely unknown. Our results showed that the targeted disruption of miR-17-92 in the testes of adult mice resulted in severe testicular atrophy, empty seminiferous tubules, and depressed sperm production. This phenotype is partly because of the reduced number of spermatogonia and spermatogonial stem cells, and the significantly increased germ cell apoptosis in the testes of miR-17-92-deficient mice. In addition, overactivation of the mammalian target of rapamycin signaling pathway and upregulation of the pro-apoptotic protein Bim, Stat3, c-Kit, and Socs3 were also observed in miR-17-92-deficient mouse testes, which might be, at least partially if not all, responsible for the aforementioned phenotypic changes in mutant testes. Taken together, these findings suggest that miR-17-92 is essential for normal spermatogenesis in mice.


Subject(s)
MicroRNAs/metabolism , Spermatogenesis/physiology , TOR Serine-Threonine Kinases/biosynthesis , Animals , Apoptosis , Apoptosis Regulatory Proteins/metabolism , Bcl-2-Like Protein 11 , Cell Line , Male , Membrane Proteins/metabolism , Mice , Proto-Oncogene Proteins/metabolism , Signal Transduction/physiology , Spermatogonia/metabolism , Testis/metabolism
12.
Oncotarget ; 6(36): 39073-87, 2015 Nov 17.
Article in English | MEDLINE | ID: mdl-26472024

ABSTRACT

Setting up breeding programs for transgenic mouse strains require to distinguish homozygous from the heterozygous transgenic animals. The combinational use of the fluorescence reporter transgene and small animal in-vivo imaging system might allow us to rapidly and visually determine the transgenic mice homozygous for transgene(s) by the in vivo fluorescence imaging. RLG, RCLG or Rm17LG transgenic mice ubiquitously express red fluorescent protein (RFP). To identify homozygous RLG transgenic mice, whole-body fluorescence imaging for all of newborn F2-generation littermates produced by mating of RFP-positive heterozygous transgenic mice (F1-generation) derived from the same transgenic founder was performed. Subsequently, the immediate data analysis of the in vivo fluorescence imaging was carried out, which greatly facilitated us to rapidly and readily distinguish RLG transgenic individual(s) with strong fluorescence from the rest of F2-generation littermates, followed by further determining this/these RLG individual(s) showing strong fluorescence to be homozygous, as strongly confirmed by mouse mating. Additionally, homozygous RCLG or Rm17LG transgenic mice were also rapidly and precisely distinguished by the above-mentioned optical approach. This approach allowed us within the shortest time period to obtain 10, 8 and 2 transgenic mice homozygous for RLG, RCLG and Rm17LG transgene, respectively, as verified by mouse mating, indicating the practicality and reliability of this optical method. Taken together, our findings fully demonstrate that the in vivo fluorescence imaging offers a visual, rapid and reliable alternative method to the traditional approaches (i.e., mouse mating and real-time quantitative PCR) in identifying homozygous transgenic mice harboring fluorescence reporter transgene under the control of a ubiquitous promoter in the situation mentioned in this study.


Subject(s)
Homozygote , Mice, Transgenic/genetics , Optical Imaging/methods , Animals , Breeding/methods , Female , Humans , Male , Mice , Mice, Inbred C57BL , Reproducibility of Results
13.
Oncotarget ; 6(34): 36713-30, 2015 Nov 03.
Article in English | MEDLINE | ID: mdl-26452025

ABSTRACT

Overexpression of the transcriptional factor Hes1 (hairy and enhancer of split-1) has been observed in numerous cancers, but the precise roles of Hes1 in epithelial-mesenchymal transition (EMT), cancer invasion and metastasis remain unknown. Our current study firstly revealed that Hes1 upregulation in a cohort of human nasopharyngeal carcinoma (NPC) biopsies is significantly associated with the EMT, invasive and metastatic phenotypes of cancer. In the present study, we found that Hes1 overexpression triggered EMT-like cellular marker alterations of NPC cells, whereas knockdown of Hes1 through shRNA reversed the EMT-like phenotypes, as strongly supported by Hes1-mediated EMT in NPC clinical specimens described above. Gain-of-function and loss-of-function experiments demonstrated that Hes1 promoted the migration and invasion of NPC cells in vitro. In addition, exogenous expression of Hes1 significantly enhanced the metastatic ability of NPC cells in vivo. Chromatin immunoprecipitation (ChIP) assays showed that Hes1 inhibited PTEN expression in NPC cells through binding to PTEN promoter region. Increased Hes1 expression and decreased PTEN expression were also observed in a cohort of NPC biopsies. Additional studies demonstrated that Hes1-induced EMT-like molecular changes and increased motility and invasion of NPC cells were mediated by PTEN. Taken together, our results suggest, for what we believe is the first time, that Hes1 plays an important role in the invasion and metastasis of NPC through inhibiting PTEN expression to trigger EMT-like phenotypes.


Subject(s)
Nasopharyngeal Neoplasms/metabolism , Nasopharyngeal Neoplasms/pathology , PTEN Phosphohydrolase/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Transcription Factor HES-1/metabolism , Animals , Carcinoma , Epithelial-Mesenchymal Transition , Female , Humans , Mice , Mice, Inbred BALB C , Mice, Nude , Middle Aged , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/genetics , Neoplasm Invasiveness , Neoplasm Metastasis , Signal Transduction , Transcription Factor HES-1/genetics
14.
Oncotarget ; 6(33): 35023-39, 2015 Oct 27.
Article in English | MEDLINE | ID: mdl-26418951

ABSTRACT

Cancer stem cells (CSCs) are considered to be the root cause for cancer treatment failure. Thus, there remains an urgent need for more potent and safer therapies against CSCs for curing cancer. In this study, the antitumor activity of cytokine-induced killer (CIK) cells against putative CSCs of nasopharyngeal carcinoma (NPC) was fully evaluated in vitro and in vivo. To visualize putative CSCs in vitro by fluorescence imaging, and image and quantify putative CSCs in tumor xenograft-bearing mice by in vivo bioluminescence imaging, NPC cells were engineered with CSC detector vector encoding GFP and luciferase (Luc) under control of Nanog promoter. Our study reported in vitro intense tumor-killing activity of CIK cells against putative CSCs of NPC, as revealed by percentage analysis of side population cells, tumorsphere formation assay and Nanog-promoter-GFP-Luc reporter gene strategy plus time-lapse recording. Additionally, time-lapse imaging firstly illustrated that GFP-labeled or PKH26-labeled putative CSCs or tumorspheres were usually attacked simultaneously by many CIK cells and finally killed by CIK cells, suggesting the necessity of achieving sufficient effector-to-target ratios. We firstly confirmed that NKG2D blockade by anti-NKG2D antibody significantly but partially abrogated CIK cell-mediated cytolysis against putative CSCs. More importantly, intravenous infusion of CIK cells significantly delayed tumor growth in NOD/SCID mice, accompanied by a remarkable reduction in putative CSC number monitored by whole-body bioluminescence imaging. Taken together, our findings suggest that CIK cells demonstrate the intense tumor-killing activity against putative CSCs of NPC, at least in part, by NKG2D-ligands recognition. These results indicate that CIK cell-based therapeutic strategy against CSCs presents a promising and safe approach for cancer treatment.


Subject(s)
Cytokine-Induced Killer Cells/transplantation , Immunotherapy, Adoptive/methods , NK Cell Lectin-Like Receptor Subfamily K/metabolism , Nasopharyngeal Neoplasms/pathology , Neoplastic Stem Cells/pathology , Animals , Blotting, Western , Carcinoma , Cell Separation , Flow Cytometry , Humans , Immunohistochemistry , In Situ Nick-End Labeling , Mice , Mice, Inbred NOD , Mice, SCID , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/metabolism , Neoplastic Stem Cells/metabolism , Transduction, Genetic , Xenograft Model Antitumor Assays
15.
Lab Invest ; 95(9): 1056-70, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26098000

ABSTRACT

The miR-19 family (miR-19a and miR-19b-1) are key oncogenic components of the miR-17-92 cluster. Overexpression of miR-19 is strongly associated with cancer invasion and metastasis, and poor prognosis of cancer patients. However, the underlying mechanisms remain largely unknown. In the present study, we found that enforced expression of miR-19 including miR-19a and miR-19b-1 triggered epithelial-mesenchymal transition (EMT) of lung cancer cells A549 and HCC827 as shown by mesenchymal-like morphological conversion, downregulation of epithelial proteins (e.g., E-cadherin, ZO-1 (zona occludens 1), and α-catenin), upregulation of mesenchymal proteins (e.g., vimentin, fibronectin 1, N-cadherin, and snail1), formation of stress fibers, and reduced cell adhesion. In addition, enhanced migration and invasion were observed in the cancer cells A549 and HCC827 undergoing EMT. In contrast, silencing of endogenous miR-19 reversed EMT and reduced the migration and invasion abilities of A549 and HCC827 cells. DNA microarray results revealed significant changes of the expression of genes related to EMT, migration, and metastasis of miR-19-expressing A549 cells. Moreover, siRNA-mediated knockdown of PTEN, a target of miR-19, also resulted in EMT, migration, and invasion of A549 and HCC827 cells, suggesting that PTEN is involved in miR-19-induced EMT, migration and invasion of lung cancer cells. Furthermore, lung cancer cells undergoing EMT induced by miR-19 demonstrated reduced proliferation in vitro and in vivo, and enhanced resistance to apoptosis caused by TNF-α. Taken together, these findings suggest that miR-19 triggers EMT, which has an important role in the invasion and migration of lung cancer cells, accompanied by the reduced proliferation of cells.


Subject(s)
Epithelial-Mesenchymal Transition/physiology , Gene Expression Regulation, Neoplastic/physiology , Lung Neoplasms/physiopathology , MicroRNAs/metabolism , Animals , Antigens, CD/metabolism , Blotting, Western , Cadherins/metabolism , Cell Adhesion/drug effects , Cell Line, Tumor , Epithelial-Mesenchymal Transition/drug effects , Fibronectins/metabolism , Gene Expression Regulation, Neoplastic/drug effects , Humans , Luciferases , Mice , Mice, Inbred BALB C , MicroRNAs/pharmacology , Oligonucleotide Array Sequence Analysis , RNA Interference , Snail Family Transcription Factors , Tetrazolium Salts , Thiazoles , Transcription Factors/metabolism , Tumor Stem Cell Assay , Vimentin/metabolism , Zonula Occludens-1 Protein/metabolism , alpha Catenin/metabolism
16.
PLoS One ; 9(7): e101330, 2014.
Article in English | MEDLINE | ID: mdl-24992599

ABSTRACT

The loss of microRNA-122 (miR-122) expression is strongly associated with increased invasion and metastasis, and poor prognosis of hepatocellular carcinoma (HCC), however, the underlying mechanisms remain poorly understood. In the present study, we observed that miR-122 over-expression in HCC cell lines Sk-hep-1 and Bel-7402 triggered the mesenchymal-epithelial transition (MET), as demonstrated by epithelial-like morphological changes, up-regulated epithelial proteins (E-cadherin, ZO-1, α-catenin, occludin, BVES, and MST4), and down-regulated mesenchymal proteins (vimentin and fibronectin). The over-expression of miRNA-122 also caused cytoskeleton disruption, RhoA/Rock pathway inactivation, enhanced cell adhesion, and suppression of migration and invasion of Sk-hep-1 and Bel-7402 cells, whereas, these effects could be reversed through miR-122 inhibition. Additional studies demonstrated that the inhibition of wild-type RhoA function induced MET and inhibited cell migration and invasion, while RhoA over-expression reversed miR-122-induced MET and inhibition of migration and invasion of HCC cells, suggesting that miR-122 induced MET and suppressed the migration and invasion of HCC cells by targeting RhoA. Moreover, our results demonstrated that HNF4α up-regulated its target gene miR-122 that subsequently induced MET and inhibited cell migration and invasion, whereas miR-122 inhibition reversed these HNF4α-induced phenotypes. These results revealed functional and mechanistic links among the tumor suppressors HNF4α, miR-122, and RhoA in EMT and invasive and metastatic phenotypes of HCC. Taken together, our study provides the first evidence that the HNF4α/miR-122/RhoA axis negatively regulates EMT and the migration and invasion of HCC cells.


Subject(s)
MicroRNAs/metabolism , rhoA GTP-Binding Protein/metabolism , 3' Untranslated Regions , Base Sequence , Cadherins/metabolism , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Cell Movement , Down-Regulation , Epithelial-Mesenchymal Transition , Hepatocyte Nuclear Factor 4/metabolism , Humans , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , MicroRNAs/antagonists & inhibitors , MicroRNAs/genetics , Oligonucleotides, Antisense/metabolism , Sequence Alignment , Signal Transduction , Transfection , Up-Regulation , Vimentin/metabolism , alpha Catenin/metabolism , rhoA GTP-Binding Protein/chemistry , rhoA GTP-Binding Protein/genetics
17.
J Asian Nat Prod Res ; 12(11): 955-61, 2010 Nov.
Article in English | MEDLINE | ID: mdl-21061217

ABSTRACT

Timosaponin BII (1), a steroidal saponin showing potential anti-dementia activity, was regioselectively hydrolyzed into its deglycosyl derivatives by the crude enzyme from Aspergillus niger AS 3.0739. Three biotransformation products, timosaponin BII-a (2), timosaponin BII-b (3), and timosaponin BII-c (4), were purified and their structures were elucidated on the basis of 1D NMR, 2D NMR, FAB-MS, and HR-ESI-MS spectral data. Compounds 2 and 3 are new compounds.


Subject(s)
Aspergillus niger/enzymology , Saponins/metabolism , Steroids/metabolism , Biotransformation , Dementia/drug therapy , Hydrolysis , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Saponins/chemistry , Saponins/pharmacology , Stereoisomerism , Steroids/chemistry , Steroids/pharmacology
18.
Carbohydr Res ; 345(12): 1752-9, 2010 Aug 16.
Article in English | MEDLINE | ID: mdl-20579977

ABSTRACT

Timosaponin BII (BII), a steroidal saponin showing potential anti-dementia activity, was converted into its glucosylation derivatives by Toruzyme 3.0L. Nine products with different degrees of glucosylation were purified and their structures were elucidated on the basis of (13)C NMR, HR-ESI-MS, and FAB-MS spectra data. The active enzyme in Toruzyme 3.0L was purified to electrophoretic homogeneity by tracking BII-glycosylase activity and was identified as Cyclodextrin-glycosyltransferase (CGTase, EC 2.4.1.19) by ESI-Q-TOF MS/MS. In this work, we found that the active enzyme catalyzed the synthesis of alpha-(1-->4)-linked glucosyl-BII when dextrin instead of an expensive activated sugar was used as the donor and showed a high thermal tolerance with the most favorable enzymatic activity at 100 degrees C. In addition, we also found that the alpha-amylases and CGTase, that is, GH13 family enzymes, all exhibited similar activities, which were able to catalyze glucosylation in steroidal saponins. But other kinds of amylases, such as gamma-amylase (GH15 family), had no such activity under the same reaction conditions.


Subject(s)
Biocatalysis , Glucosyltransferases/metabolism , Saponins/metabolism , Steroids/metabolism , Carbohydrate Sequence , Enzyme Activation , Glucosyltransferases/chemistry , Glycosylation , Hydrogen-Ion Concentration , Molecular Conformation , Saponins/chemistry , Stereoisomerism , Steroids/chemistry , Temperature , Time Factors
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