RESUMO
This study investigated the expression of microRNA-210 (miR-210) in tissue and serum of renal carcinoma patients and its effect on renal carcinoma cell proliferation, apoptosis, and invasion. Thirty-two renal carcinoma patients in our hospital were selected as the study group and 32 people receiving a physical examination were selected as the control group. miR-210 expression in the serum of renal carcinoma patients and in healthy subjects was quantified by real-time polymerase chain reaction. After miR-210 overexpression and inhibition in ACHN cells in human renal carcinoma, ACHN cell proliferation, apoptosis, and invasion were detected by CCK-8, flow cytometry, and a transwell invasion assay. The expression of miR-210 was significantly higher in renal carcinoma than in corresponding paracarcinoma tissues (P < 0.001). The expression of miR-210 was significantly higher in the serum of renal carcinoma than in the control group (P < 0.001). ACHN cell proliferation and invasion were significantly increased and apoptosis was significantly decreased (P < 0.05) when miR-210 was overexpressed. ACHN cell proliferation and invasion were significantly decreased and apoptosis was significantly increased (P < 0.05) when miR-210 was inhibited. In conclusion, miR-210 was highly expressed in tissues and serum of renal carcinoma patients. miR-210 could promote the proliferation and invasion of renal carcinoma cells and inhibit the apoptosis of renal cell carcinoma cells.
Assuntos
Apoptose , Carcinoma de Células Renais/metabolismo , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Neoplasias Renais/metabolismo , MicroRNAs/genética , Invasividade Neoplásica , Idoso , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/patologia , Feminino , Humanos , Neoplasias Renais/genética , Neoplasias Renais/patologia , Masculino , Pessoa de Meia-Idade , Regulação para CimaRESUMO
Transforming growth factor beta 1 (TGF-ß1) and bone morphogenetic protein-2 (BMP-2) are important regulators of bone repair and regeneration. In this study, we examined whether TGF-ß1 and BMP-2 expressions were delayed during bone healing in type 1 diabetes mellitus. Tibial fractures were created in 95 diabetic and 95 control adult male Wistar rats of 10 weeks of age. At 1, 2, 3, 4, and 5 weeks after fracture induction, five rats were sacrificed from each group. The expressions of TGF-ß1 and BMP2 in the fractured tibias were measured by immunohistochemistry and quantitative reverse-transcription polymerase chain reaction, weekly for the first 5 weeks post-fracture. Mechanical parameters (bending rigidity, torsional rigidity, destruction torque) of the healing bones were also assessed at 3, 4, and 5 weeks post-fracture, after the rats were sacrificed. The bending rigidity, torsional rigidity and destruction torque of the two groups increased continuously during the healing process. The diabetes group had lower mean values for bending rigidity, torsional rigidity and destruction torque compared with the control group (P<0.05). TGF-ß1 and BMP-2 expression were significantly lower (P<0.05) in the control group than in the diabetes group at postoperative weeks 1, 2, and 3. Peak levels of TGF-ß1 and BMP-2 expression were delayed by 1 week in the diabetes group compared with the control group. Our results demonstrate that there was a delayed recovery in the biomechanical function of the fractured bones in diabetic rats. This delay may be associated with a delayed expression of the growth factors TGF-ß1 and BMP-2.
Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Calo Ósseo/fisiopatologia , Diabetes Mellitus Tipo 1/fisiopatologia , Consolidação da Fratura/fisiologia , Fraturas da Tíbia/fisiopatologia , Fator de Crescimento Transformador beta1/metabolismo , Animais , Fenômenos Biomecânicos , Diabetes Mellitus Experimental/fisiopatologia , Diabetes Mellitus Tipo 1/metabolismo , Fraturas Ósseas/fisiopatologia , Imuno-Histoquímica , Masculino , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Fraturas da Tíbia/metabolismo , Fatores de Tempo , TorqueRESUMO
Transforming growth factor beta 1 (TGF-β1) and bone morphogenetic protein-2 (BMP-2) are important regulators of bone repair and regeneration. In this study, we examined whether TGF-β1 and BMP-2 expressions were delayed during bone healing in type 1 diabetes mellitus. Tibial fractures were created in 95 diabetic and 95 control adult male Wistar rats of 10 weeks of age. At 1, 2, 3, 4, and 5 weeks after fracture induction, five rats were sacrificed from each group. The expressions of TGF-β1 and BMP2 in the fractured tibias were measured by immunohistochemistry and quantitative reverse-transcription polymerase chain reaction, weekly for the first 5 weeks post-fracture. Mechanical parameters (bending rigidity, torsional rigidity, destruction torque) of the healing bones were also assessed at 3, 4, and 5 weeks post-fracture, after the rats were sacrificed. The bending rigidity, torsional rigidity and destruction torque of the two groups increased continuously during the healing process. The diabetes group had lower mean values for bending rigidity, torsional rigidity and destruction torque compared with the control group (P<0.05). TGF-β1 and BMP-2 expression were significantly lower (P<0.05) in the control group than in the diabetes group at postoperative weeks 1, 2, and 3. Peak levels of TGF-β1 and BMP-2 expression were delayed by 1 week in the diabetes group compared with the control group. Our results demonstrate that there was a delayed recovery in the biomechanical function of the fractured bones in diabetic rats. This delay may be associated with a delayed expression of the growth factors TGF-β1 and BMP-2.
Assuntos
Animais , Masculino , Fraturas da Tíbia/fisiopatologia , Calo Ósseo/fisiopatologia , Consolidação da Fratura/fisiologia , Diabetes Mellitus Tipo 1/fisiopatologia , Fator de Crescimento Transformador beta1/metabolismo , Proteína Morfogenética Óssea 2/metabolismo , Fraturas da Tíbia/metabolismo , Fatores de Tempo , Fenômenos Biomecânicos , Imuno-Histoquímica , Ratos Wistar , Torque , Diabetes Mellitus Experimental/fisiopatologia , Diabetes Mellitus Tipo 1/metabolismo , Fraturas Ósseas/fisiopatologia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
The morphological species concept is based on morpho-logical traits, which are often subject to subjectivity or artifact. Molecular evidence is needed to test the reliability of morphological classification of taxa that are controversial and to provide appropriate taxonomic de-limitation. In this study, we used 15 single-copy nuclear loci and 2 chloroplast fragments to verify the morphological classification of the Salix matsudana Koidz. complex using phylogenetic approaches. Complete sequence alignment showed slight diversification in nuclear sequences and no variety in chloroplast DNA fragments. Phylogenetic trees revealed a monophyletic group consisting of all individuals of S. matsudana and 2 clades within this group, with a 100% bootstrap support value and 1.00 posterior probability. The topology of the phylogenetic trees was highly consistent with the morphological classification of the S. matsudana complex. Verifying the genetic background of these classification units based on remarkable morphological differences will provide a foundation for future studies of Salix and the breeding of new horticultural varieties.
Assuntos
Salix/genética , Genes de Plantas , Estudos de Associação Genética , Loci Gênicos , Tipagem Molecular , Filogenia , Proteínas de Plantas/genética , Salix/anatomia & histologia , Salix/classificação , Análise de Sequência de DNARESUMO
The expression of transforming growth factor-beta 1 (TGF-ß1) inside the callus cells of diabetic rats and the impact of insulin therapy on its expression and biomechanics was investigated. The rats were randomly divided as follows: an insulin therapy group (IT), a diabetic model group (DM), and a non-diabetic control group (NC). Bone specimens from each group were extracted at different times for immunohistochemical observation of the expression of TGF-ß1. Concurrently, the destruction torque and torsional stiffness were detected at different times. One to four weeks after fracture, TGF-ß1 was widely expressed in fractured callus cells and periosteal proliferating cells, while the expression inside diabetic cells was significantly reduced. The expression of TGF-ß1 decreased over the first 68 weeks, and the mature bone cells never expressed TGF-ß1. The destruction torque (Nm) detected in the 6th week revealed that there was a statistically significant difference between the DM, NC, and IT groups (P < 0.01). In conclusion, TGF-ß1 expression was significantly reduced inside the callus cells of diabetic rats. Insulin therapy increased TGF-ß1 expression inside the callus cells of diabetic rats and improved the biomechanical characteristics of the callus.
Assuntos
Calo Ósseo/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Hipoglicemiantes/farmacologia , Insulina/farmacologia , Fraturas da Tíbia/tratamento farmacológico , Fator de Crescimento Transformador beta1/genética , Animais , Calo Ósseo/metabolismo , Calo Ósseo/patologia , Proliferação de Células , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Consolidação da Fratura/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Dureza , Masculino , Periósteo/efeitos dos fármacos , Periósteo/metabolismo , Periósteo/patologia , Ratos , Ratos Wistar , Fraturas da Tíbia/complicações , Fraturas da Tíbia/genética , Fraturas da Tíbia/patologia , Torque , Fator de Crescimento Transformador beta1/metabolismoRESUMO
We explored whether p53 upregulated modulator of apoptosis (PUMA) gene transfection could enhance the sensitivity of epirubicin-induced apoptosis of MCF-7 breast cancer cells. The liposome-mediated recombinant eukaryotic expression vector PU-MA-pCDNA3 and empty vector plasmid were stably transfected into MCF-7 cells. Epirubicin (0.01-100 µM) was applied to MCF-7, MCF-7/PUMA, and MCF-7/pCDNA3 cells for 72 h. The MTT assay was used to calculate the cell survival rate in each group, and the 50% inhibitory concentration (IC50) was calculated. The IC50 values of epirubicin in MCF-7, MCF-7/PUMA, and MCF-7/pCDNA3 cells were 13 ± 1.4, 1.8 ± 0.2, and 10.7 ± 1.3 µM, respectively. The sensitivity of MCF-7/PUMA cells to epirubicin increased 7.2-fold. Epirubicin induced apoptosis in MCF-7 cells dose-dependently, but MCF-7/PUMA cell-induced apoptosis was more significant compared to controls. Low concentrations of epirubicin (0.1 µM) caused low levels of apoptosis of MCF-7/pCDNA3 (1.15 ± 0.26%) and MCF-7 cells (0.9 ± 0.24%), but significantly induced apoptosis of MCF-7/PUMA cells (6.44 ± 1.46%). High epirubicin concentration (1 µM) induced apoptosis in each group, but the epirubicin MCF-7/PUMA apoptosis rate (35.47 ± 9.36%) was significantly higher than that of MCF-7 (12.6 ± 3.73%) and MCF-7/ pCDNA3 (15.2 ± 5.17%) cells (P < 0 01). Flow cytometry and TUNEL assays for apoptosis detection showed similar results. PUMA protein expression in MCF-7/PUMA cells was significantly higher than that in MCF-7 and MCF-7/pCDNA3 cells by Western blot analysis. There-fore, stable transfection of PUMA can significantly enhance epirubicin-induced apoptosis sensitivity of MCF-7 breast cancer cells.
Assuntos
Proteínas Reguladoras de Apoptose/genética , Apoptose/genética , Neoplasias da Mama/genética , Proliferação de Células/genética , Proteínas Proto-Oncogênicas/genética , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/biossíntese , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Proliferação de Células/efeitos dos fármacos , Epirubicina/administração & dosagem , Feminino , Citometria de Fluxo , Regulação Neoplásica da Expressão Gênica , Humanos , Células MCF-7 , Proteínas Proto-Oncogênicas/biossínteseRESUMO
Increasing evidence shows that TIM-1 and TIM-3 in-fluence chronic autoimmune diseases, and their expression levels in immune cells from nephritic patients are still unknown. Real-time transcription-polymerase chain reaction analysis was used to deter-mine expression levels of TIM-1 and TIM-3 mRNA in peripheral blood mononuclear cells (PBMCs) from 36 patients with minimal change glo-merulopathy (MCG), 65 patients with lupus nephritis (LN), 78 patients with IgA nephropathy (IgAN), 55 patients with membranous nephropa-thy (MN), 22 patients with crescentic glomerulonephritis (CGN), 26 patients with anaphylactoid purpura nephritis (APN), and 63 healthy controls. TIM-3 mRNA expression significantly decreased in PBMCs from nephritic patients (LN, P < 0.0001; MCG, P < 0.0001; MN, P = 0.0031; CGN, P = 0.0464; IgAN, P = 0.0002; APN, P = 0.0392) com-pared with healthy controls. In contrast, there was no significant differ-ence in TIM-1 mRNA expression between the patients and the healthy controls. Our results suggest that insufficient expression of TIM-3 mRNA may be involved in the pathogenesis of nephropathy.
Assuntos
Glomerulonefrite por IGA/sangue , Nefrite Lúpica/sangue , Glicoproteínas de Membrana/biossíntese , Proteínas de Membrana/biossíntese , Receptores Virais/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Regulação Neoplásica da Expressão Gênica , Glomerulonefrite por IGA/genética , Glomerulonefrite por IGA/patologia , Receptor Celular 1 do Vírus da Hepatite A , Receptor Celular 2 do Vírus da Hepatite A , Humanos , Leucócitos Mononucleares , Nefrite Lúpica/genética , Nefrite Lúpica/patologia , Masculino , Glicoproteínas de Membrana/sangue , Proteínas de Membrana/sangue , Pessoa de Meia-Idade , RNA Mensageiro/biossíntese , RNA Mensageiro/sangue , Receptores Virais/sangueRESUMO
The T-cell immunoglobulin and mucin domain (TIM) family is associated with autoimmune diseases, but its expression level in the immune cells of systemic lupus erythematosus (SLE) patients is not known. The aim of this study was to investigate whether the expression of TIM-3 mRNA is associated with pathogenesis of SLE. Quantitative real-time reverse transcription-polymerase chain reaction analysis (qRT-PCR) was used to determine TIM-1, TIM-3, and TIM-4 mRNA expression in peripheral blood mononuclear cells (PBMCs) from 132 patients with SLE and 62 healthy controls. The PBMC surface protein expression of TIMs in PBMCs from 20 SLE patients and 15 healthy controls was assayed by flow cytometry. Only TIM-3 mRNA expression decreased significantly in SLE patients compared with healthy controls (P<0.001). No significant differences in TIM family protein expression were observed in leukocytes from SLE patients and healthy controls (P>0.05). SLE patients with lupus nephritis (LN) had a significantly lower expression of TIM-3 mRNA than those without LN (P=0.001). There was no significant difference in the expression of TIM-3 mRNA within different classes of LN (P>0.05). Correlation of TIM-3 mRNA expression with serum IgA was highly significant (r=0.425, P=0.004), but was weakly correlated with total serum protein (rs=0.283, P=0.049) and serum albumin (rs=0.297, P=0.047). TIM-3 mRNA expression was weakly correlated with the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI; rs=-0.272, P=0.032). Our results suggest that below-normal expression of TIM-3 mRNA in PBMC may be involved in the pathogenesis of SLE.
RESUMO
The T-cell immunoglobulin and mucin domain (TIM) family is associated with autoimmune diseases, but its expression level in the immune cells of systemic lupus erythematosus (SLE) patients is not known. The aim of this study was to investigate whether the expression of TIM-3 mRNA is associated with pathogenesis of SLE. Quantitative real-time reverse transcription-polymerase chain reaction analysis (qRT-PCR) was used to determine TIM-1, TIM-3, and TIM-4 mRNA expression in peripheral blood mononuclear cells (PBMCs) from 132 patients with SLE and 62 healthy controls. The PBMC surface protein expression of TIMs in PBMCs from 20 SLE patients and 15 healthy controls was assayed by flow cytometry. Only TIM-3 mRNA expression decreased significantly in SLE patients compared with healthy controls (P<0.001). No significant differences in TIM family protein expression were observed in leukocytes from SLE patients and healthy controls (P>0.05). SLE patients with lupus nephritis (LN) had a significantly lower expression of TIM-3 mRNA than those without LN (P=0.001). There was no significant difference in the expression of TIM-3 mRNA within different classes of LN (P>0.05). Correlation of TIM-3 mRNA expression with serum IgA was highly significant (r=0.425, P=0.004), but was weakly correlated with total serum protein (rs=0.283, P=0.049) and serum albumin (rs=0.297, P=0.047). TIM-3 mRNA expression was weakly correlated with the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI; rs=-0.272, P=0.032). Our results suggest that below-normal expression of TIM-3 mRNA in PBMC may be involved in the pathogenesis of SLE.
RESUMO
Species of Populus are widely distributed worldwide, playing a significant role in both ecology and economy. However, the lack of single-copy nuclear markers limits knowledge about the phylogeny and population genetics of this genus. In the present study, primer pairs of 15 single-copy nuclear markers were developed through bioinformatic methods based on complete genomic sequences of Populus trichocarpa and Salix arbutifolia. Twenty individuals of Populus davidiana Dode and Salix matsudana Koidz were used to evaluate the basic application of these markers with respect to marker length and diversity indices, respectively. The utility of single-copy nuclear markers is anticipated to facilitate further studies about the phylogeny, population genetics, and phylogeography of this genus, in addition to providing information about the evolutionary dynamics of Salicaceae.