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Functional analysis of microRNAs in human hepatocellular cancer stem cells.

Meng F, Glaser SS, Francis H, DeMorrow S, Han Y, Passarini JD, Stokes A, Cleary JP, Liu X, Venter J, Kumar P, Priester S, Hubble L, Staloch D, Sharma J, Liu CG, Alpini G - J. Cell. Mol. Med. (2012)

Bottom Line: Importantly, inhibition of let-7 increases the chemosensitivity of HSCs to sorafenib and doxorubicin whereas silencing of miR-181 led to a reduction in HSCs motility and invasion.Knocking down IL-6 and Twist in HSCs significantly reduced let-7 and miR-181 expression and subsequently inhibited chemoresistance and cell invasion.In conclusion, alterations of IL-6- and Twist-regulated microRNA expression in HSCs play a part in tumour spreading and responsiveness to chemotherapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, TX 76504, USA.

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miRNA expression profiles in human HCC cancer stem cells [under self-renewal (SR) or differentiation (DF)], malignant and non-malignant hepatocytes. (A) miRNA was isolated and profiling by hybridization to miRNA-specific probes on epoxy-coated slides. Cluster analysis identifies a group of miRNA that are increased in expression in HCC cancer stem cells (HSC-SR and HSC-DF) and reduced in hepG2 cells when compared to normal human liver stem cells (Cluster 2). An enlarged view of this group, containing 11 miRNAs including members of let-7 and miR-181 family is shown. Lower middle panel illustrated representative Northern blot analysis of let-7a and miR-181a using total RNA isolated from four cell lines. This cluster of miRNAs are among the group, which increased in HSC-SR by fourfold, and P < 0.05 when compared to HepG2 cells (lower right panel). (B) The expressions of mature let-7a, let-7b and miR-181a miRNAs were validated using real-time PCR and correlated with the pattern in the miRNA array. Data represent mean ± S.E. from four separate experiments.
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fig03: miRNA expression profiles in human HCC cancer stem cells [under self-renewal (SR) or differentiation (DF)], malignant and non-malignant hepatocytes. (A) miRNA was isolated and profiling by hybridization to miRNA-specific probes on epoxy-coated slides. Cluster analysis identifies a group of miRNA that are increased in expression in HCC cancer stem cells (HSC-SR and HSC-DF) and reduced in hepG2 cells when compared to normal human liver stem cells (Cluster 2). An enlarged view of this group, containing 11 miRNAs including members of let-7 and miR-181 family is shown. Lower middle panel illustrated representative Northern blot analysis of let-7a and miR-181a using total RNA isolated from four cell lines. This cluster of miRNAs are among the group, which increased in HSC-SR by fourfold, and P < 0.05 when compared to HepG2 cells (lower right panel). (B) The expressions of mature let-7a, let-7b and miR-181a miRNAs were validated using real-time PCR and correlated with the pattern in the miRNA array. Data represent mean ± S.E. from four separate experiments.

Mentions: We then compared miRNA expression between HCC CSCs and HepG2/N-LSCs. The pattern of miRNA expression in HCC CSCs was markedly different from that of either normal human liver stem cells or the HepG2 HCC cells (Fig. 3A). The expression of a group of miRNA was differentially increased in HSC cells. These include the human let-7 and miR-181 family members, all of which were expressed >4.0-fold in HSC-SR and HSC-DF CSCs compared with HepG2 HCC cells as well as normal liver stem cells (Fig. 3B). Aberrant expressions of let-7a, let-7b and miR-181a were verified by Northern blot (Fig. 3A) as well as real-time PCR analysis (Fig. 3B). Furthermore, silencing IL-6 and Twist in HSCs significantly reduced let-7a and miR-181a expression respectfully, but not vice versa (Fig. S2), implicated the specific link between IL-6 and let-7, as well as Twist and miR-181 in tumour stem cell behaviour.


Functional analysis of microRNAs in human hepatocellular cancer stem cells.

Meng F, Glaser SS, Francis H, DeMorrow S, Han Y, Passarini JD, Stokes A, Cleary JP, Liu X, Venter J, Kumar P, Priester S, Hubble L, Staloch D, Sharma J, Liu CG, Alpini G - J. Cell. Mol. Med. (2012)

miRNA expression profiles in human HCC cancer stem cells [under self-renewal (SR) or differentiation (DF)], malignant and non-malignant hepatocytes. (A) miRNA was isolated and profiling by hybridization to miRNA-specific probes on epoxy-coated slides. Cluster analysis identifies a group of miRNA that are increased in expression in HCC cancer stem cells (HSC-SR and HSC-DF) and reduced in hepG2 cells when compared to normal human liver stem cells (Cluster 2). An enlarged view of this group, containing 11 miRNAs including members of let-7 and miR-181 family is shown. Lower middle panel illustrated representative Northern blot analysis of let-7a and miR-181a using total RNA isolated from four cell lines. This cluster of miRNAs are among the group, which increased in HSC-SR by fourfold, and P < 0.05 when compared to HepG2 cells (lower right panel). (B) The expressions of mature let-7a, let-7b and miR-181a miRNAs were validated using real-time PCR and correlated with the pattern in the miRNA array. Data represent mean ± S.E. from four separate experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3116063&req=5

fig03: miRNA expression profiles in human HCC cancer stem cells [under self-renewal (SR) or differentiation (DF)], malignant and non-malignant hepatocytes. (A) miRNA was isolated and profiling by hybridization to miRNA-specific probes on epoxy-coated slides. Cluster analysis identifies a group of miRNA that are increased in expression in HCC cancer stem cells (HSC-SR and HSC-DF) and reduced in hepG2 cells when compared to normal human liver stem cells (Cluster 2). An enlarged view of this group, containing 11 miRNAs including members of let-7 and miR-181 family is shown. Lower middle panel illustrated representative Northern blot analysis of let-7a and miR-181a using total RNA isolated from four cell lines. This cluster of miRNAs are among the group, which increased in HSC-SR by fourfold, and P < 0.05 when compared to HepG2 cells (lower right panel). (B) The expressions of mature let-7a, let-7b and miR-181a miRNAs were validated using real-time PCR and correlated with the pattern in the miRNA array. Data represent mean ± S.E. from four separate experiments.
Mentions: We then compared miRNA expression between HCC CSCs and HepG2/N-LSCs. The pattern of miRNA expression in HCC CSCs was markedly different from that of either normal human liver stem cells or the HepG2 HCC cells (Fig. 3A). The expression of a group of miRNA was differentially increased in HSC cells. These include the human let-7 and miR-181 family members, all of which were expressed >4.0-fold in HSC-SR and HSC-DF CSCs compared with HepG2 HCC cells as well as normal liver stem cells (Fig. 3B). Aberrant expressions of let-7a, let-7b and miR-181a were verified by Northern blot (Fig. 3A) as well as real-time PCR analysis (Fig. 3B). Furthermore, silencing IL-6 and Twist in HSCs significantly reduced let-7a and miR-181a expression respectfully, but not vice versa (Fig. S2), implicated the specific link between IL-6 and let-7, as well as Twist and miR-181 in tumour stem cell behaviour.

Bottom Line: Importantly, inhibition of let-7 increases the chemosensitivity of HSCs to sorafenib and doxorubicin whereas silencing of miR-181 led to a reduction in HSCs motility and invasion.Knocking down IL-6 and Twist in HSCs significantly reduced let-7 and miR-181 expression and subsequently inhibited chemoresistance and cell invasion.In conclusion, alterations of IL-6- and Twist-regulated microRNA expression in HSCs play a part in tumour spreading and responsiveness to chemotherapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, TX 76504, USA.

Show MeSH
Related in: MedlinePlus