ABSTRACT
ObjectiveTo explore the role of long non-coding RNA (lncRNA)-H19 in the proliferation and migration of non-small cell lung cancer (NSCLC) cells and the molecular mechanisms. MethodsThe expressions of lncRNA-H19 in 20 NSCLC tissues and paired non-tumor tissues, which were collected from Changhai Hospital of Naval Medical University (Second Military Medical University) from Oct. 2015 to May 2016, were detected by real-time quantitative PCR (qPCR). We also examined lncRNA-H19 expressions in NSCLC cell lines A549 and NCI-H1299 and normal lung epithelial cell line BEAS-2B by qPCR. The proliferation and migration of A549 cells overexpressing lncRNA-H19 were detected by CCK-8 assay and Transwell assay, respectively. Bioinformatics analysis and duel-luciferase reporter assay were conducted to predict and confirm the interaction between microRNA (miRNA)-760 and lncRNA-H19. Western blotting analysis and RT-qPCR were performed to observe the influence of lncRNA-H19 overexpression on the expression of miRNA-760 and target gene nanog. MiRNA-760 was overexpressed in A549 cells, and its role in lncRNA-H19 promoting proliferation and migration of NSCLC cells was observed. Results The expressions of lncRNA-H19 in NSCLC tissues and A549 and NCI-H1299 cells were significantly upregulated compared with those in normal tissues and BEAS-2B cells (all P<0.01). Overexpression of lncRNA-H19 significantly improved the proliferation ability (P<0.05) and migration ability (P<0.01) of A549 cells compared with the negative control group. The results of starBase v3.0 showed that lncRNA-H19 could specifically adsorb miRNA-760, and duel-luciferase reporter assay showed that lncRNA-H19 directly bound to miRNA-760. Compared with the negative control group, overexpression of lncRNA-H19 significantly inhibited miRNA-760 expression in A549 cells (P<0.05) and promoted the expression of the downstream gene nanog at mRNA and protein levels (all P<0.01). Overexpression of miRNA-760 significantly inhibited lncRNA-H19-induced proliferation and migration of A549 cells (all P <0.05). ConclusionLncRNA-H19 can promote the proliferation and migration of NSCLC cells through sponging miRNA-760 to regulate nanog gene expression.
ABSTRACT
Objective To investigate the expression of NANOG gene in cervical cancer and its effect on the development of the cancer. Methods By using immunohistochemistry, NANOG expression levels were demonstrated in normal cervical tissues, cervical cancer in situ and cervical cancer tissues. Tumor sphere cells from cervical cancer tissue and their differentiated cells were injected into nude mice to observe the tumor growth. Results When compared with that of the normal cervical tissue, significantly higher NANOG protein expression level was found in cervical cancer and cervical cancer in situ (P<0.05). Tumor spheres were formed from the non-serum culture of cervical cancer tissue and the spheres could differentiate into epithelial-like cells. The sphere cells of cervical cancer caused tumor formation in nude mice. Conclusion The results suggest that cancer stem cells are present in cervical cancer and NANOG may play a crucial role in the development of cervical cancer.
ABSTRACT
Las autorrenovación y la diferenciación son características de las células madre que varían entre los diferentes tipos celulares según el tejido en el que se encuentren y el microambiente que las rodee. En ambos procesos intervienen inhibidores del ciclo celular, genes implicados en rearreglos cromosómicos, proteínas del desarrollo esencial y vías de señalización específicas. La autorrenovación está regulada por diversos mecanismos, entre los cuales se destacan las vías Wnt, Notch y Hedgehog, y los factores BMI-1, p16Ink4a, ARF, NANOG, OCT3/4, SOX2, HOXB4 y sus páralogos. Los adelantos en el conocimiento de la biología de las células madre y de los mecanismos moleculares que regulan la autorrenovación y la diferenciación han convertido a estas células en una importante promesa para la investigación básica y aplicada.
Self-renewal capacity and differentiation are features of stem cells that vary among the different cellular types according to the tissue in which they reside and the surroundingmicroenvironment. Cellular cycle inhibitors, genes implied in chromosomal rearrangements, essential development proteins and specific signaling pathways intervene in these processes. Self-renewal is regulated by different mechanisms, the most important of which are the Wnt, Notch and Hedgehogpathways, and the factors BMI-1, p16Ink4a, ARF, NANOG, OCT3/4, SOX2, HOXB4 and their paralogs. Advancesin the knowledge of stem cells biology and of the molecular mechanisms that influence their selfrenewal and differentiation have made these cellsan important promise for both basic and applied research.
Subject(s)
Stem Cells , Cell Differentiation , ADP-Ribosylation Factor 1 , Transcription Factors/physiologyABSTRACT
In order to further study mouse embryonic stem cells(ES cells),lentiviral vector PLL-IRES-Nanog-Neo was constructed.Mouse ES cells overexpressed nanog by mediation of lentiviral were cultured on mouse fetal fibroblast feeders after 2 weeks under G418 media and examined according to gowth characteristics. Results were showed that 918 bp nanog fragments were expressed in mouse ES cells mediated by lentiviral vector PLL-IRES-Nanog-Neo,mouse nanog-ES cells were taken on mass-like image and positve with alkaline phosphatase staining and Oct4 and SSEA1 immunocytochemistry under no LIF condition in the media. It is concluded that mouse ES cells Elevated nanog gene expression by mediation of lentiviral were constucted and cultured.