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Our previous study demonstrated that human KIAA0100 gene is a novel acute monocytic leukemia-associated antigen (MLAA) gene. But the functional characterization of human KIAA0100 gene has remained unknown to date. Here, firstly, bioinformatic prediction of human KIAA0100 gene was carried out using online software;Secondly, human KIAA0100 gene expression was downregulated by the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system in U937 cells. Cell proliferation and apoptosis were next evaluated in KIAA0100-knockdown U937 cells. The bioinformatic prediction showed that human KIAA0100 gene was located on 17q11.2, and human KIAA0100 protein was located in the secretory pathway. Besides, human KIAA0100 protein contained a signal peptide, a transmembrane region, three types of secondary structures (alpha helix, extended strand, and random coil) , and four domains from mitochondrial protein 27 (FMP27). The observation on functional characterization of human KIAA0100 gene revealed that its downregulation inhibited cell proliferation, and promoted cell apoptosis in U937 cells. To summarize, these results suggest human KIAA0100 gene possibly comes within mitochondrial genome; moreover, it is a novel anti-apoptotic factor related to carcinogenesis or progression in acute monocytic leukemia, and may be a potential target for immunotherapy against acute monocytic leukemia.
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BACKGROUND:Inhibiting the apoptosis of intervertebral disc cel s can postpone the degenerative process of intervertebral disc. Survivin has a strong function of regulating cel proliferation and anti-apoptosis. OBJECTIVE:To construct and identify the lentiviral vector encoding survivin gene of human. METHODS:The survivin gene of human (BIRC5) was synthesized through the gene synthesis technology, amplified by PCR and analyzed by electrophoresis. The target gene was cloned into lentiviral expression plasmid to obtain the recombinant lentiviral vector Lenti-BIRC5. After transformation into competent E. coli cel s, the candidate clones were identified by PCR firstly. The positive clones were identified by gene sequencing. The lentivirus plasmid containing target gene was transfected into 293T cel s, and the expression of recombinant lentiviral vector Flag-Survivin fusion protein was detected through western blot analysis. RESULTS AND CONCLUSION:The PCR results of electrophoresis and DNA sequencing showed that lentiviral vector containing human survivin gene was constructed successful y. Western blot analysis results showed that the target gene was transfected successful y and over-expressed in cultured cel s. The lentiviral expression vector of human survivin gene Lenti-BIRC5 was constructed successful y, which lays a foundation for the study addressing the anti-apoptotic effects of survivin on human nucleus pulposus cel s.
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BACKGROUND:Insulin-like growth factor 1 is the key factor during cartilage development, which is involved in the growth and reconstruction of condylar cartilage. OBJECTIVE:To study the effect of insulin-like growth factor 1 on cel apoptosis and the apopotosis-associated factors of Bcl-2, Bax mRNA and protein expressions of rat condylar chondrocytes. METHODS:The 1-day-old and 28-day-old rat condylar chondrocytes were cultured and identified in vitro. The condylar chondrocytes with different ages were divided into experimental group and control group. After being starved for 24 hours, chondrocytes in the experimental group were incubated with 100μg/L recombined rat insulin-like growth factor 1 for 48 hours, while the chondrocytes in the control group were incubated normal y. RESULTS AND CONCLUSION:Compared with the control group, after being incubated with recombined insulin-like growth factor 1, the number of condylar chondrocytes was increased with high speed proliferation (Pproliferation and reduce cel apoptosis of newborn and adolescent rat condylar chondrocytes, which may be mediated by Bcl-2 and Bax.