RESUMO
The aim of this study was to investigate the expression of T-cell immunoglobulin mucin domain molecule-3 (Tim-3) in osteosarcoma tissues, and analyze its effect on cell proliferation and metastasis in an osteosarcoma cell line. Tim-3 mRNA and protein expression in osteosarcoma tissue was detected by reverse transcriptase-polymerase chain reaction and immunohistochemistry, respectively. Additionally, the cell viability, apoptosis rate, and invasive ability of the osteosarcoma cell line MG-63 were tested using the methyl thiazolyl tetrazolium assay, Annexin V-propidium iodide flow cytometry, and a Transwell assay, respectively, following Tim-3 interference using small interfering RNA (siRNA). We also analyzed the expression of Snail, E-cadherin, vimentin, and nuclear factor (NF)-kB in the cells by western blot. We observed that Tim-3 mRNA and protein was significantly overexpressed in osteosarcoma tissues, compared to the adjacent normal tissue (P < 0.01). Moreover, MG-63 cells transfected with the Tim-3 siRNA presented lower cell viability, a greater number of apoptotic cells, and decreased invasive ability (P < 0.01), compared to control cells. Additionally, we observed a decrease in Snail and vimentin expression, an increase in the E-cadherin level, and an increase in NF-kB p65 phosphorylation (P < 0.01) in Tim-3 siRNA-transfected MG-63 cells. Based on these results, we concluded that Tim-3 is highly expressed in osteosarcoma tissue. Moreover, we speculated that interfering in Tim-3 expression could significantly suppress osteosarcoma cell (MG-63) proliferation and metastasis via the NF-kB/Snail signaling pathway and epithelial-mesenchymal transition.
Assuntos
Neoplasias Ósseas/genética , Transição Epitelial-Mesenquimal/genética , Regulação Neoplásica da Expressão Gênica , Receptor Celular 2 do Vírus da Hepatite A/genética , Osteossarcoma/genética , Antígenos CD , Apoptose , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Caderinas/genética , Caderinas/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Colágeno/química , Cultura em Câmaras de Difusão , Combinação de Medicamentos , Receptor Celular 2 do Vírus da Hepatite A/antagonistas & inibidores , Receptor Celular 2 do Vírus da Hepatite A/metabolismo , Humanos , Laminina/química , Metástase Linfática , Invasividade Neoplásica , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Fosforilação , Proteoglicanas/química , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Fatores de Transcrição da Família Snail/genética , Fatores de Transcrição da Família Snail/metabolismo , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Vimentina/genética , Vimentina/metabolismoRESUMO
This study aims to explore the roles of somatic embryogenesis receptor-like kinase (SERK) in Malus hupehensis (Pingyi Tiancha). The full-length sequences of SERK1 in triploid Pingyi Tiancha (3n) and a tetraploid hybrid strain 33# (4n) were cloned, sequenced, and designated as MhSERK1 and MhdSERK1, respectively. Multiple alignments of amino acid sequences were conducted to identify similarity between MhSERK1 and MhdSERK1 and SERK sequences in other species, and a neighbor-joining phylogenetic tree was constructed to elucidate their phylogenetic relations. Expression levels of MhSERK1 and MhdSERK1 in different tissues and developmental stages were investigated using quantitative real-time PCR. The coding sequence lengths of MhSERK1 and MhdSERK1 were 1899 bp (encoding 632 amino acids) and 1881 bp (encoding 626 amino acids), respectively. Sequence analysis demonstrated that MhSERK1 and MhdSERK1 display high similarity to SERKs in other species, with a conserved intron/exon structure that is unique to members of the SERK family. Additionally, the phylogenetic tree showed that MhSERK1 and MhdSERK1 clustered with orange CitSERK (93%). Furthermore, MhSERK1 and MhdSERK1 were mainly expressed in the reproductive organs, in particular the ovary. Their expression levels were highest in young flowers and they differed among different tissues and organs. Our results suggest that MhSERK1 and MhdSERK1 are related to plant reproduction, and that MhSERK1 is related to apomixis in triploid Pingyi Tiancha.
Assuntos
Flores/genética , Malus/genética , Filogenia , Sequência de Aminoácidos/genética , Proteínas de Arabidopsis/genética , Clonagem Molecular , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Malus/crescimento & desenvolvimento , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Proteínas Quinases/genética , Alinhamento de Sequência , Tetraploidia , TriploidiaRESUMO
Serial data from studies of infants at the University of Iowa and from the Fels Longitudinal Study were used to develop sex-specific percentiles for increments in weight and recumbent length for selected intervals during the first 24 months of life. Weight increments are presented for 1-month intervals from birth to 6 months, 2-month intervals from birth to 12 months, and 3-month intervals from birth to 24 months. Length increments are presented for 2-month intervals from birth to 6 months, and for 3-month intervals from birth to 24 months of age. Weights and lengths at the target ages were obtained for the Iowa data by simple interpolation, and for the Fels data by fitting families of three-parameter mathematical functions to the serial data from ages 1 to 24 months. The tabular presentations are based on the Iowa data from birth to 3 months of age, on the combined Iowa and Fels data from 3 to 6 months of age, and on the Fels data from 6 to 24 months of age. We believe that these reference data will be useful in screening for deviations from normal growth and may aid in early detection of failure to thrive or excessive weight gain during early life.