iBet uBet web content aggregator. Adding the entire web to your favor.
iBet uBet web content aggregator. Adding the entire web to your favor.



Link to original content: http://www.ncbi.nlm.nih.gov/pubmed/25202400
Downregulation of GSK3β by miR-544a to maintain self-renewal ability of lung caner stem cells - PubMed 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.

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.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Oct;8(4):1731-1734.
doi: 10.3892/ol.2014.2387. Epub 2014 Jul 28.

Downregulation of GSK3β by miR-544a to maintain self-renewal ability of lung caner stem cells

Xiao-Mei Mo  1 , Hua-Hui Li  2 , Ming Liu  3 , Yan-Tuan Li  3
Affiliations

Downregulation of GSK3β by miR-544a to maintain self-renewal ability of lung caner stem cells

Xiao-Mei Mo et al. Oncol Lett. 2014 Oct.

Abstract

In order to study the influence and mechanism of miR-544a on the self-renewal ability of lung cancer stem cells, TargetScan was used to predict the target gene of miR-544a. A luciferase reporter system and western blotting were used to validate the target genes identified by TargetScan. 95C and 95D low and high metastatic human lung cancer cells were transfected with miR-544a, and quantitative polymerase chain reaction (qPCR) was used to verify the miR-544a expression in these two cell lines. Tumor ball (spheroid) suspension culture was use to study the effects of miR-544a on lung cancer stem cells. TargetScan predicted that miR-544a interacted with GSK3β. A luciferase reporter system (F=201.37, P<0.01) and western blot analysis was used to validate that miR-544a could inhibit the expression of GSK3β, while β-catenin and CD133 were significantly increased in miR-544a-overexpressing 95C and 95D cells (F=9.43, 7.73 and 3.37, respectively; P<0.01). qPCR revealed that miR-544a was overexpressed in transfected 95C and 95D cells (20.51±0.97 and 15.16±1.38, respectively; F=418.05; P<0.01). miR-544a-overexpressing cells formed spheroids in suspension cultures of spheroid single cells. miR-544a was shown to reduce the expression of GSK3β and activate the Wnt signaling pathway to maintain the self-renewal ability of lung caner stem cells.

Keywords: GSK3β; Wnt signaling pathway; cancer stem cells; miR-544a; non-small cell lung cancer.

PubMed Disclaimer

Figures

Figure 1
Figure 1
miR-544a can combine with the 3′UTR of GSK3β. miR, microRNA; 3′UTR, 3′ untranslated region.
Figure 2
Figure 2
Western blot analysis of protein expression in the Wnt pathway. NC, negative control; mimc, mimic combine; miR, microRNA; 95C, low metastatic human lung cancer cells; 95D, high metastatic human lung cancer cells.
Figure 3
Figure 3
Impact of miR-544a in spheroid formation (magnification, ×200). NC, negative control; mir, microRNA; 95C, low metastatic human lung cancer cells; 95D, high metastatic human lung cancer cells.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Lam WK, Watkins DN. Lung cancer: future directions. Respirology. 2007;12:471–477. - PubMed
    1. Lobo NA, Shimono Y, Qian D, et al. The biology of cancer stem cells. Annu Rev Cell Dev Biol. 2007;23:675–699. - PubMed
    1. Dean M, Fojo T, Bates S. Tumor stem cells and drug resistance. Nat Rev Cancer. 2005;5:275–284. - PubMed
    1. Lin CT, Lyu YL, Xiao H, et al. Suppression of gene amplification and chromosomal DNA integration by the DNA mismatch repair system. Nucleic Acids Res. 2001;29:3304–3310. - PMC - PubMed
    1. Alamgeer M, Peacock CD, Matsui W, et al. Cancer stem cells in lung cancer: Evidence and controversies. Respirology. 2013;18:757–764. - PMC - PubMed