Objective: To elucidate the relation between human tissue factor pathwayinhibitor-2 (TFPI-2) expression and ovarian tumor migration and invasion. Methods: Human TFPI-2expression vector pBos-Cite-neo/TFPI-2 was transfe...Objective: To elucidate the relation between human tissue factor pathwayinhibitor-2 (TFPI-2) expression and ovarian tumor migration and invasion. Methods: Human TFPI-2expression vector pBos-Cite-neo/TFPI-2 was transfected into ovarian tumor cells line A2780- Afterthe transfected cells were selected by G418, transfected and nontransfected cells were screened forTFPI-2 mRNA and protein by reverse transcription-polymerase chain reaction and Western blotanalysis, respectively. The number of transfected or nontransfected cells passing through membraneof Boyden chamber was counted as the basis assessing tumor cells migratory and invasive behaviors.Results: Expression of mRNA and protein of TFPI-2 was detectable in transfected cells. In invasionassay, the number of TFPI-2-expressing cells to traverse a Matrigel-coated membrane was obviouslydecreased compared with that of nonexpressing cells (59.3±6.5 vs 109.7±5.5, P 【 0.01); While inmigration assay, no significant difference through a noncoated membrane was observed amongtransfected and nontransfected cells (114.7±8.6 vs 127.3±7.1, P 】 0.05). Conclusion: Expression ofTFPI-2 may strongly inhibit the invasive ability of ovarian tumor cells in vitro, but has no effecton the migratory ability which provides an experimental basis for genotherapy of human ovariantumor.展开更多
MicroRNAs(miRNAs) are a group of small, endogenous, single-stranded non-coding RNAs that post-transcriptionally regulate gene expression levels. Previous studies have revealed that miRNAs play key roles in multiple bi...MicroRNAs(miRNAs) are a group of small, endogenous, single-stranded non-coding RNAs that post-transcriptionally regulate gene expression levels. Previous studies have revealed that miRNAs play key roles in multiple biological processes, such as growth and development in both animals and plants. Computational identification is an efficient method for miRNA prediction in organisms with a reference genome before high-throughput miRNA sequencing experiments. In this study, we employed an integrated strategy combining the homology-based and ab initio approaches to predict miRNAs from the genome and transcriptome of large yellow croaker, one of the most commercially important marine fish in China and East Asia. A total of 418 miRNA molecules, including 287 miRNAs by the homology-based method and 131 miRNAs by the ab initio approach, were identified for large yellow croaker. Additionally, 16 053 target genes were predicted and annotated for Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) databases. Meanwhile, we analysed single nucleotide polymorphisms(SNPs) around large yellow croaker miRNA and found that the miRNA seed regions were significantly less prone to mutations, indicating that the miRNA sequences were under strict natural selection based on their essential regulation functions in living cells. Twenty-two SNPs were identified in large yellow croaker miRNA seed regions, which dramatically influenced the miRNA-gene regulation networks. This is the first reported miRNA detection from both the genome and transcriptome using the integrated strategy for large yellow croaker species, and the miRNA and SNP analyses in this work provide important resources and a reference for subsequent miRNA functional investigations in large yellow croaker.展开更多
文摘Objective: To elucidate the relation between human tissue factor pathwayinhibitor-2 (TFPI-2) expression and ovarian tumor migration and invasion. Methods: Human TFPI-2expression vector pBos-Cite-neo/TFPI-2 was transfected into ovarian tumor cells line A2780- Afterthe transfected cells were selected by G418, transfected and nontransfected cells were screened forTFPI-2 mRNA and protein by reverse transcription-polymerase chain reaction and Western blotanalysis, respectively. The number of transfected or nontransfected cells passing through membraneof Boyden chamber was counted as the basis assessing tumor cells migratory and invasive behaviors.Results: Expression of mRNA and protein of TFPI-2 was detectable in transfected cells. In invasionassay, the number of TFPI-2-expressing cells to traverse a Matrigel-coated membrane was obviouslydecreased compared with that of nonexpressing cells (59.3±6.5 vs 109.7±5.5, P 【 0.01); While inmigration assay, no significant difference through a noncoated membrane was observed amongtransfected and nontransfected cells (114.7±8.6 vs 127.3±7.1, P 】 0.05). Conclusion: Expression ofTFPI-2 may strongly inhibit the invasive ability of ovarian tumor cells in vitro, but has no effecton the migratory ability which provides an experimental basis for genotherapy of human ovariantumor.
基金Supported by the National Natural Science Foundation of China(Nos.U1205122,31602207)the Key Project of the Xiamen Southern Ocean Research Center(No.14GZY70NF34)+2 种基金the Fujian Provincial Natural Science Foundation(No.2016J05081)the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A403)the Foundation for Innovation Research Team of Jimei University(No.2010A02)
文摘MicroRNAs(miRNAs) are a group of small, endogenous, single-stranded non-coding RNAs that post-transcriptionally regulate gene expression levels. Previous studies have revealed that miRNAs play key roles in multiple biological processes, such as growth and development in both animals and plants. Computational identification is an efficient method for miRNA prediction in organisms with a reference genome before high-throughput miRNA sequencing experiments. In this study, we employed an integrated strategy combining the homology-based and ab initio approaches to predict miRNAs from the genome and transcriptome of large yellow croaker, one of the most commercially important marine fish in China and East Asia. A total of 418 miRNA molecules, including 287 miRNAs by the homology-based method and 131 miRNAs by the ab initio approach, were identified for large yellow croaker. Additionally, 16 053 target genes were predicted and annotated for Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) databases. Meanwhile, we analysed single nucleotide polymorphisms(SNPs) around large yellow croaker miRNA and found that the miRNA seed regions were significantly less prone to mutations, indicating that the miRNA sequences were under strict natural selection based on their essential regulation functions in living cells. Twenty-two SNPs were identified in large yellow croaker miRNA seed regions, which dramatically influenced the miRNA-gene regulation networks. This is the first reported miRNA detection from both the genome and transcriptome using the integrated strategy for large yellow croaker species, and the miRNA and SNP analyses in this work provide important resources and a reference for subsequent miRNA functional investigations in large yellow croaker.