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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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Twist is involved in cell motility and invasion potentials if HCC stem cells. (A) Light microscopy was used to document morphologic differences between the Twist and control siRNA treated N-LSC, HepG2 and HSCs. HSCs displayed an aggressive phenotype, which became elongated, and forming the leading edge pseudopodia. Silencing Twist in HSCs significantly reduced the percentage of cells with invasive phenotypes. (B) Matrigel invasion assay showing the invading capability of normal and malignant liver stem cells with and without Twist silencing. (C, D) Cell migration and invasion was assessed in Oct-4+CD133+ and Oct-4−CD133− cells with or without Twist silencing. Cell migration was expressed as arbitrary fluorescence units. Cell invasion was assessed in 96-well plates pre-coated with extracellular matrix. RFU is the absolute florescence intensity value and FI is the florescence intensity after normalization with the standard curve. The mean and standard error from four separate experiments are illustrated. #P < 0.05 when compare with HepG2 and N-LSC groups. *P < 0.05 when compared with siRNA controls.
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fig07: Twist is involved in cell motility and invasion potentials if HCC stem cells. (A) Light microscopy was used to document morphologic differences between the Twist and control siRNA treated N-LSC, HepG2 and HSCs. HSCs displayed an aggressive phenotype, which became elongated, and forming the leading edge pseudopodia. Silencing Twist in HSCs significantly reduced the percentage of cells with invasive phenotypes. (B) Matrigel invasion assay showing the invading capability of normal and malignant liver stem cells with and without Twist silencing. (C, D) Cell migration and invasion was assessed in Oct-4+CD133+ and Oct-4−CD133− cells with or without Twist silencing. Cell migration was expressed as arbitrary fluorescence units. Cell invasion was assessed in 96-well plates pre-coated with extracellular matrix. RFU is the absolute florescence intensity value and FI is the florescence intensity after normalization with the standard curve. The mean and standard error from four separate experiments are illustrated. #P < 0.05 when compare with HepG2 and N-LSC groups. *P < 0.05 when compared with siRNA controls.

Mentions: It has been suggested that tumour invasion and metastasis may be mediated by the CSC population [43, 44]. We observed human HCC cells and stem cells by light microscopy to examine invasion of the Matrigel. Movements of malignant cells in Matrigel were associated with formation of peripheral ruffles. Compared to HepG2 and N-LSC cells, HSCs had more cells with membrane ruffles, and cells were more invasive in the Matrigel (Fig. 7A and B). In vitro assays were used to compare sorted subpopulations from the perspective of differences in cell migration, and invasion (Fig. 7C and D). Oct-4+CD133+ cells were observed to be significantly more migratory towards serum (Fig. 6C), and significantly more invasive through ECM gel (Fig. 7D) than respective Oct-4−CD133− cell subsets (P < 0.001).


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)

Twist is involved in cell motility and invasion potentials if HCC stem cells. (A) Light microscopy was used to document morphologic differences between the Twist and control siRNA treated N-LSC, HepG2 and HSCs. HSCs displayed an aggressive phenotype, which became elongated, and forming the leading edge pseudopodia. Silencing Twist in HSCs significantly reduced the percentage of cells with invasive phenotypes. (B) Matrigel invasion assay showing the invading capability of normal and malignant liver stem cells with and without Twist silencing. (C, D) Cell migration and invasion was assessed in Oct-4+CD133+ and Oct-4−CD133− cells with or without Twist silencing. Cell migration was expressed as arbitrary fluorescence units. Cell invasion was assessed in 96-well plates pre-coated with extracellular matrix. RFU is the absolute florescence intensity value and FI is the florescence intensity after normalization with the standard curve. The mean and standard error from four separate experiments are illustrated. #P < 0.05 when compare with HepG2 and N-LSC groups. *P < 0.05 when compared with siRNA controls.
© Copyright Policy
Related In: Results  -  Collection

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

fig07: Twist is involved in cell motility and invasion potentials if HCC stem cells. (A) Light microscopy was used to document morphologic differences between the Twist and control siRNA treated N-LSC, HepG2 and HSCs. HSCs displayed an aggressive phenotype, which became elongated, and forming the leading edge pseudopodia. Silencing Twist in HSCs significantly reduced the percentage of cells with invasive phenotypes. (B) Matrigel invasion assay showing the invading capability of normal and malignant liver stem cells with and without Twist silencing. (C, D) Cell migration and invasion was assessed in Oct-4+CD133+ and Oct-4−CD133− cells with or without Twist silencing. Cell migration was expressed as arbitrary fluorescence units. Cell invasion was assessed in 96-well plates pre-coated with extracellular matrix. RFU is the absolute florescence intensity value and FI is the florescence intensity after normalization with the standard curve. The mean and standard error from four separate experiments are illustrated. #P < 0.05 when compare with HepG2 and N-LSC groups. *P < 0.05 when compared with siRNA controls.
Mentions: It has been suggested that tumour invasion and metastasis may be mediated by the CSC population [43, 44]. We observed human HCC cells and stem cells by light microscopy to examine invasion of the Matrigel. Movements of malignant cells in Matrigel were associated with formation of peripheral ruffles. Compared to HepG2 and N-LSC cells, HSCs had more cells with membrane ruffles, and cells were more invasive in the Matrigel (Fig. 7A and B). In vitro assays were used to compare sorted subpopulations from the perspective of differences in cell migration, and invasion (Fig. 7C and D). Oct-4+CD133+ cells were observed to be significantly more migratory towards serum (Fig. 6C), and significantly more invasive through ECM gel (Fig. 7D) than respective Oct-4−CD133− cell subsets (P < 0.001).

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