Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or . mil. Before sharing sensitive information, make sure you’re on a federal government site. VSports app下载.

Https

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely. V体育官网.

. 2019 Feb 13;20(4):801.
doi: 10.3390/ijms20040801.

Long Non Coding RNA H19: A New Player in Hypoxia-Induced Multiple Myeloma Cell Dissemination

Chiara Corrado  1 , Viviana Costa  2 , Gianluca Giavaresi  3   4 , Annalisa Calabrese  5 , Alice Conigliaro  6 , Riccardo Alessandro  7
Affiliations

"VSports在线直播" Long Non Coding RNA H19: A New Player in Hypoxia-Induced Multiple Myeloma Cell Dissemination

Chiara Corrado et al. Int J Mol Sci. .

Abstract

The long non-coding RNA H19 (lncH19) is broadly transcribed in the first stage of development and silenced in most cells of an adult organism; it appears again in several tumors where, through different molecular mediators, promotes cell proliferation, motility and metastases. LncH19 has been associated with hypoxia-inducible factor 1-alpha (HIF-1α) activation and, in some tumors, it has proved to be necessary and required to sustain hypoxic responses. Here we propose to investigate a putative role for the lncH19 in hypoxia induced multiple myeloma (MM) progression. Transcriptional analysis of MM cell lines (RPMI and MM1 VSports手机版. S) exposed to normoxia or hypoxia (1% O₂) was done in order to evaluate lncH19 levels under hypoxic stimulation. Then, to investigate the role of lncH19 in hypoxia mediated MM progression, transcriptional, protein and functional assays have been performed on hypoxia stimulated MM cell lines, silenced or not for lncH19. Our data demonstrated that hypoxic stimulation in MM cell lines induced the overexpression of lncH19, which, in turn, is required for the expression of the hypoxia induced genes involved in MM dissemination, such as C-X-C Motif Chemokine Receptor 4 (CXCR4) and Snail. Moreover, adhesion assays demonstrated that lncH19 silencing abrogates the increased adhesion on stromal cells induced by the hypoxic condition. Finally, Western blot analysis indicated that lncH19 silencing impaired HIF1α nuclear translocation. The LncH19, required for the induction of hypoxic responses in MM cells, could represent a new therapeutic target for MM. .

Keywords: HIF-1α; hypoxia; long non-coding RNA H19 (lncH19); multiple myeloma V体育安卓版. .

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure A1
Figure A1
ELISA assay for nuclear HIF1-α indicates that three different multiple myeloma cell lines respond to 24-h hypoxic chamber stimulation. Values are presented as mean ± SD. Statistical analysis was performed by Student-t test: * p < 0.05; ** p < 0.01.
Figure 1
Figure 1
qRT-PCR indicate long non coding H19 (lncH19) levels in Multiple Myeloma (MM) cell lines after 24-h hypoxic stimulation expressed as fold of induction versus lncH19 levels in normoxia. Statistical analysis was performed by the use of student t-test; * p < 0.05; ** p < 0.01 (A). qRT-PCR indicate the basal level of the lncH19 in normoxic MM cell lines. Statistical analysis was performed by the use of one way ANOVA test and Dunnett’s multiple comparison test; *** p = 0.001 (B). qRT-PCR indicate the levels of miR-675-5p in MM cell lines after 24-h hypoxic stimulation expressed as fold of induction versus normoxia (C). Values are presented as mean ± SD.
Figure 2
Figure 2
qRT-PCR indicate the H19 expression levels after hypoxic stimulation in MM cell lines infected with siH19 and relative controls. Value are expressed as Fold Of Increase (FOI) respect to siRNA Scramble (siScr) infected cells (A). qRT-PCR of indicated genes in MM cell lines after hypoxic stimulation compared to normoxia. Value are expressed as FOI respect to normoxic cells (B). qRT-PCR of indicated genes in hypoxic MM cell lines silenced or not for lncH19. Value are expressed as FOI respect to siScr infected cells (C). Values are presented as mean ± SD. Statistical analysis was performed by the use of Student t-test. * p < 0.05; ** p < 0.001; *** p < 0.0001.
Figure 3
Figure 3
Adhesion assay of MM cells to stromal monolayer, in normoxia and after hypoxic stimulation, silenced or not for lncH19. Representative images of different experimental condition captured by Nikon A1 confocal microscope, scale bar = 50 μm (A); quantification of Green Fluorescent Protein (GFP) positive MM adherent cells. Values are presented as mean ± SD. Statistical analysis was performed by the use of one way ANOVA test and Dunnett’s multiple comparison test ** p < 0.001; *** p < 0.0001 (B).
Figure 4
Figure 4
qRT-PCR (A) and Western blot analysis on total protein extract (B) of HIF-1α expression in MM cells silenced or not for lncH19. Densitometric analysis with Image J software was done with respect to total protein level of β-actin, used as loading control. (C) Western blot analysis on nuclear extract of MM hypoxic cells silenced or not for lncH19. Densitometric analysis with Image J software was done with respect to nuclear protein level of histone H3, used as loading control. Values are presented as mean ± SD.
Figure 5
Figure 5
Western blot analysis of IPO7 and VHL on total extract of MM hypoxic cells silenced, or not, for lncH19. Densitometric analysis with Image J software was done with respect to total protein level of β-actin, used as loading control. Values are presented as mean ± SD.
See this image and copyright information in PMC

V体育安卓版 - References

    1. Diamantopoulos M.A., Tsiakanikas P., Scorilas A. Non-coding RNAs: The riddle of the transcriptome and their perspectives in cancer. Ann. Transl. Med. 2018;6:241. doi: 10.21037/atm.2018.06.10. - "V体育2025版" DOI - PMC - PubMed
    1. Sun H., Huang Z., Sheng W., Xu M.D. Emerging roles of long non-coding RNAs in tumor metabolism. J. Hematol. Oncol. 2018;11:106. doi: 10.1186/s13045-018-0648-7. - DOI - PMC - PubMed
    1. Raveh E., Matouk I.J., Gilon M., Hochberg A. The H19 Long non-coding RNA in cancer initiation, progression and metastasis—A proposed unifying theory. Mol. Cancer. 2015;14:184. doi: 10.1186/s12943-015-0458-2. - DOI - PMC - PubMed
    1. Yoshimura H., Matsuda Y., Yamamoto M., Kamiya S., Ishiwata T. Expression and role of long non-coding RNA H19 in carcinogenesis. Front. Biosci. (Landmark Ed.) 2018;23:614–625. - PubMed
    1. Zhou J., Yang L., Zhong T., Mueller M., Men Y., Zhang N., Xie J., Giang K., Chung H., Sun X., et al. H19 lncRNA alters DNA methylation genome wide by regulating S-adenosylhomocysteine hydrolase. Nat. Commun. 2015;6:10221. doi: 10.1038/ncomms10221. - DOI - PMC - PubMed

Substances (VSports app下载)