ABSTRACT
OBJECTIVE: To screen the genes that may play an important role in the carcinogenesis of nasopharyngeal carcinoma (NPC). METHODS: Microdissection and cDNA genechip hybridization techniques were used to examine the differentially expressed genes in NPC tissue, the surrounding and adjacent tissues of NPC, and the nasopharyngeal inflammation tissue. The fluorescent signals on cDNA chip were scanned and the results of hybridization analyzed by image processing software. RESULTS: Many differentially expressed genes were identified between the three samples, including many different types of genes, such as those responsible for signal and protein transmission, oncogene and tumor suppression genes, immune-associated genes, apoptosis genes and DNA binding and transcription factor genes. CONCLUSION: The carcinogenesis of NPC involves many genes of a variety of types, suggesting its complex process.
Subject(s)
Carcinoma, Squamous Cell/genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Nasopharyngeal Neoplasms/genetics , Carcinoma, Squamous Cell/pathology , DNA, Complementary/genetics , DNA, Neoplasm/genetics , Female , Humans , Male , Nasopharyngeal Neoplasms/pathology , Neoplasm Invasiveness , Oligonucleotide Array Sequence AnalysisABSTRACT
The GATA-1 of Xenopus (xGATA-1), which has two subtypes xGATA-1a and xGATA-1b, is a necessary factor for erythroid differentiation and maturation as similar as that of other GATA-1s. Although both xGATa-1a and xGATA-1b are able to stimulate erythropoiesis, only xGATA-1b is capable of inhibiting neurogenesis in Xenopus embryos. Compared between their structures, xGATA-1a and xGATA-1b are very similar in nucleotide and amino acids composition, but not identical. Therefore, it is responsible for studying the role of the diverse codons between the two genes, so the desired mutations: S(168), H(169) double deletion and point mutation of T(304)-->A, T(359)-->A, were introduced into xGATA-1b gene through site-directed mutagenesis. Then, mRNA from each mutant as well as wtxGATA-1b was co-injected with DN-BR mRNA or separately injected into Xenopus stage 2 embryos, and the role of mutants in erythropoiesis and neurogenesis was analyzed by using animal cap culture system. The results showed that the neural-inhibiting activity of xGATA-1b, but not hematopoiesis-inducing activity, was aborted because of deletion of Ser(168) and His(169) or point mutation of T(359)-->A. So it is demonstrated for the first time that Ser(168) and His(169) or Thr(359)in xGATA-1b may be one of the structural basis for explanting the different function between xGATA-1b and xGATA-1a.
Subject(s)
Amino Acids/genetics , DNA-Binding Proteins/genetics , Mutagenesis, Site-Directed , Transcription Factors/genetics , Animals , DNA-Binding Proteins/physiology , Embryo, Nonmammalian/drug effects , Embryo, Nonmammalian/metabolism , Embryonic Development , Erythroid-Specific DNA-Binding Factors , Erythropoiesis/physiology , Histidine/genetics , Injections , Mutation , Nervous System/embryology , Protein Isoforms/genetics , Protein Isoforms/physiology , RNA, Messenger/administration & dosage , RNA, Messenger/genetics , RNA, Messenger/metabolism , Serine/genetics , Threonine/genetics , Transcription Factors/physiology , Xenopus laevisABSTRACT
BACKGROUND & OBJECTIVE: The mechanism of how stromal cell play an important role in nasopharyngeal carcinogenesis is now a hotspot. This study was designed to elucidate the possible mechanism of stromal cell in the occurrence and progression of nasopharyngeal carcinoma (NPC) through the analysis of the characteristics of gene expression in pericancerous stromal cells of NPC by cDNA array. METHODS: The atlas human select tumor arrays were used to compare the expression profiles between NPC tissue and NPC cell lines. RESULTS: Pericancerous stromal cells in NPC expressed at least 40 genes specifically. CONCLUSION: The specific expression of these genes in pericancerous stromal cells provides energy materials for growth of NPC cells; furthermore, it can accelerate the degradation of extracellular matrix, thus promoting the metastasis of NPC cells.
Subject(s)
DNA, Neoplasm/analysis , Gene Expression , Nasopharyngeal Neoplasms/genetics , Stromal Cells/physiology , DNA, Complementary/analysis , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Humans , Oligonucleotide Array Sequence Analysis/methodsABSTRACT
BACKGROUND & OBJECTIVE: Microdissection has become indispensable for the selective analysis of stroma-free tumor cell. However, To obtain sufficient RNA from microdissected cells is difficult. The study was designed to seek a specific way to separate nasopharyngeal carcinoma (NPC) cells from stromal cells and to amplify the RNA from microdissected NPC cells. METHODS: NPC cells were obtained using microdissection from frozen NPC tissue sections, then RNA was extracted from the microdissected NPC cells and reverse transcribed in vitro. The expression levels of beta-actin and GADPH in amplified RNA were detected using RT-PCR. RESULTS: About 20,000-40,000 NPC cells were obtained, RNA was extracted from the cells, about 0.5-2.5 kb RNA fragments were obtained after RNA linear amplification and beta-actin and GADPH levels were integral. CONCLUSION: Microdissection combined with RNA linear amplification can be used to successfully obtain pure NPC cells, the integrity of amplified RNA is good and can be used in further research.