ABSTRACT
BACKGROUND:The functions of homo heterogeneous ribonucleoprotein E1 are very wide. It can participate in the expression of skeleton proteins in the nervous system. OBJECTIVE:To construct the recombinant plasmid of homo heterogeneous ribonucleoprotein E1 and observe its expression in nerve cells for further studying the functions of it in neurocytes. METHODS:Using pcDNATM4/His C, the homo heterogeneous ribonucleoprotein E1 was subcloned into recombinant plasmid E1-pcDNATM 4/His C, fol owed by enzyming and sequencing. After SH-SY5Y cells were transfected with the recombinant plasmid, western blot analysis and real time RT-PCR were used to detect the expression of homo heterogeneous ribonucleoprotein E1 in SH-SY5Y cells. And the growth of SH-SY5Y cells was observed. RESULTS AND CONCLUSION:We successful y constructed the eukaryotic expressed vector of homo heterogeneous ribonucleoprotein E1. The recombinant plasmids were verified to express in SH-SY5Y cells correctly at mRNA and protein levels. And SH-SY5Y cells generated quickly after homo heterogeneous ribonucleoprotein E1 was over-expressed. The homo heterogeneous ribonucleoprotein E1 is an important protein in neural development. And this vector offers the premise for further studying its function in nervous system.
ABSTRACT
Objective: PCBP1 is a family member of heterogeneous nuclear ribonucleoproteins (hnRNPs) that belong to RNA-binding proteins and bear three KH domains. The protein plays a pivotal role in post-transcriptional regulation for RNA metabolism and RNA function in gene expression. We hy-pothesized and were going to identify that the regulatory function of PCBP1 is performed through different complexes of proteins that include PCBP1. Methods: To test our hypothesis, approaches of protein wal-king with a yeast two-hybrid system (Y2H), pulling down in yeasts, co-immunoprecipitation and immu-nofluorescent microscopy assay were employed in this study. The PCBP1 was used as the initial "walker" to search for its interaction partner(s). Results: Candidate proteins including MYL6, PECAM1, CSH1,RAB7, p57KIP2, ACTG1, RBMS1 and PSG4-1ike were identified with selection mediums and preceding methods. Conclusion: With these candidate protein molecules, some protein complexes associating with PCBP1 are proposed, which may help in a better understanding of physiological functions of PCBP1 and proved evidence that PCBP1 is involved in variant biological pathways.
ABSTRACT
BACKGROUND: Cytogenetic evidence suggests that chromosomal alteration are not randomly occurring events and some malignancies are characterized by specific chromosome abnormalities, which provides cytogenetic basis for the expression of oncogenes.OBJECTIVE: To investigate the characteristics of chromosomal karyotype and marker chromosome of breast cancer cell lines Bcap-37and MCF-7 by means of G-banding chromosomal analysis.DESIGN: A controlled experiment with breast cancer cells as observation subjects.SETTING: Department of Medical Genetics, Peking University Health Science Center.MATERIALS: This study was carried out in the Department of Medical Genetics of Peking University Health Science Center from April 1991 to May 1992 using breast cancer cell lines MCF-7 and Bcap-37.METHODS: The chromosomes of human breast cancer cell lines Bcap-37and MCF-7 were obtained by growth synchronization induced by hypothermia and colchicines treatment. The cells at prometaphase or metaphase underwent G-banding chromosomal analysis. For each cell line, 50 to 60 mitotic figures were counted and 15 or 16 G-binding karyotypes were analyzed, including the mitotic figure at the level of about 320- and 500-band .MAIN OUTCOME MEASURES:Abnormality in chromosome number and structural aberration of the two breast cancer cell lines.RESULTS:The modal chromosomal number of Bcap-37 cell line was 63, of which 17 marker chromosomes had identifiable structure, as compared with 13 out of 56 chromosomes in modal number of MCF-7 cell line.CONCLUSION: Both of the two breast cancer cell lines have complex cytogenetic abnormality in the modal number and structure of the chromosomes, which might result in the rearrangement of DNA sequence of the cancer-related genes or DNA depletion, so as to play important roles in the pathogenesis and development of breast cancer.