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Downregulation of long non-coding RNA H19 promotes P19CL6 cells proliferation and inhibits apoptosis during late-stage cardiac differentiation via miR-19b-modulated Sox6

View Article: PubMed Central - PubMed

ABSTRACT

Background: Regulating cardiac differentiation to maintain normal heart development and function is very important. At present, biological functions of H19 in cardiac differentiation is not completely clear.

Methods: To explore the functional effect of H19 during cardiac differentiation. Expression levels of early cardiac-specific markers Nkx-2.5 and GATA4, cardiac contractile protein genes α-MHC and MLC-2v were determined by qRT-PCR and western lot. The levels of lncRNA H19 and miR-19b were detected by qRT-PCR. We further predicted the binding sequence of H19 and miR-19b by online softwares starBase v2.0 and TargetScan. The biological functions of H19 and Sox6 were evaluated by CCK-8 kit, cell cycle and apoptosis assay and caspase-3 activity.

Results: The expression levels of α-MHC, MLC-2v and H19 were upregulated, and miR-19b was downregulated significantly in mouse P19CL6 cells at the late stage of cardiac differentiation. Biological function analysis showed that knockdown of H19 promoted cell proliferation and inhibits cell apoptosis. H19 suppressed miR-19b expression and miR-19b targeted Sox6, which inhibited cell proliferation and promoted apoptosis in P19CL6 cells during late-stage cardiac differentiation. Importantly, Sox6 overexpression could reverse the positive effects of H19 knockdown on P19CL6 cells.

Conclusion: Downregulation of H19 promoted cell proliferation and inhibited cell apoptosis during late-stage cardiac differentiation by regulating the negative role of miR-19b in Sox6 expression, which suggested that the manipulation of H19 expression could serve as a potential strategy for heart disease.

No MeSH data available.


Related in: MedlinePlus

Sox6 reversed the proliferation and antiapoptosis effects of H19 knockdown on P19CL6 cells. a CCK-8 assay indicated shH19 boosted the viability of P19CL6 cells at day 8 and day 10, and pcDNA-Sox6 reversed the effect. b Cell cycle assay found that shH19 increased the percentage of cells in the S phase at day 8 and 10, whereas pcDNA-Sox6 attenuated the effect. (c and d) The rate of apoptosis and caspase-3 activity were significantly decreased in cells treated with shH19 at day 8 and 10, and pcDNA-Sox6 overturned the effect. *P < 0.05, **P < 0.01, ***P < 0.001
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Fig6: Sox6 reversed the proliferation and antiapoptosis effects of H19 knockdown on P19CL6 cells. a CCK-8 assay indicated shH19 boosted the viability of P19CL6 cells at day 8 and day 10, and pcDNA-Sox6 reversed the effect. b Cell cycle assay found that shH19 increased the percentage of cells in the S phase at day 8 and 10, whereas pcDNA-Sox6 attenuated the effect. (c and d) The rate of apoptosis and caspase-3 activity were significantly decreased in cells treated with shH19 at day 8 and 10, and pcDNA-Sox6 overturned the effect. *P < 0.05, **P < 0.01, ***P < 0.001

Mentions: To further testify the link between H19 and Sox6, we transfected shH19 or shH19 + pcDNA-Sox6 into P19CL6 cells to investigate their effects on cell proliferation and apoptosis. CCK-8 assay indicated that shH19 significantly promoted the viability of P19CL6 cells at day 6, 8 and 10, but pcDNA-Sox6 attenuated the effect of shH19 on cell viability (Fig. 6a). Cell cycle assay indicated that shH19 markedly increased the percentage of cells in (S + G2/M)/(G0 + G1), while pcDNA-Sox6 overturned the effect (Fig. 6b). Futhermore, we found that the rate of apoptosis and caspase-3 activity were obviously reduced in cells treated with shH19, however, pcDNA-Sox6 reversed the anti-apoptosis effect of shH19 on P19CL6 cells (Fig. 6c and d). All of the data suggested that H19 regulated P19CL6 cell proliferation and apoptosis by modulating Sox6 expression.Fig. 6


Downregulation of long non-coding RNA H19 promotes P19CL6 cells proliferation and inhibits apoptosis during late-stage cardiac differentiation via miR-19b-modulated Sox6
Sox6 reversed the proliferation and antiapoptosis effects of H19 knockdown on P19CL6 cells. a CCK-8 assay indicated shH19 boosted the viability of P19CL6 cells at day 8 and day 10, and pcDNA-Sox6 reversed the effect. b Cell cycle assay found that shH19 increased the percentage of cells in the S phase at day 8 and 10, whereas pcDNA-Sox6 attenuated the effect. (c and d) The rate of apoptosis and caspase-3 activity were significantly decreased in cells treated with shH19 at day 8 and 10, and pcDNA-Sox6 overturned the effect. *P < 0.05, **P < 0.01, ***P < 0.001
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Fig6: Sox6 reversed the proliferation and antiapoptosis effects of H19 knockdown on P19CL6 cells. a CCK-8 assay indicated shH19 boosted the viability of P19CL6 cells at day 8 and day 10, and pcDNA-Sox6 reversed the effect. b Cell cycle assay found that shH19 increased the percentage of cells in the S phase at day 8 and 10, whereas pcDNA-Sox6 attenuated the effect. (c and d) The rate of apoptosis and caspase-3 activity were significantly decreased in cells treated with shH19 at day 8 and 10, and pcDNA-Sox6 overturned the effect. *P < 0.05, **P < 0.01, ***P < 0.001
Mentions: To further testify the link between H19 and Sox6, we transfected shH19 or shH19 + pcDNA-Sox6 into P19CL6 cells to investigate their effects on cell proliferation and apoptosis. CCK-8 assay indicated that shH19 significantly promoted the viability of P19CL6 cells at day 6, 8 and 10, but pcDNA-Sox6 attenuated the effect of shH19 on cell viability (Fig. 6a). Cell cycle assay indicated that shH19 markedly increased the percentage of cells in (S + G2/M)/(G0 + G1), while pcDNA-Sox6 overturned the effect (Fig. 6b). Futhermore, we found that the rate of apoptosis and caspase-3 activity were obviously reduced in cells treated with shH19, however, pcDNA-Sox6 reversed the anti-apoptosis effect of shH19 on P19CL6 cells (Fig. 6c and d). All of the data suggested that H19 regulated P19CL6 cell proliferation and apoptosis by modulating Sox6 expression.Fig. 6

View Article: PubMed Central - PubMed

ABSTRACT

Background: Regulating cardiac differentiation to maintain normal heart development and function is very important. At present, biological functions of H19 in cardiac differentiation is not completely clear.

Methods: To explore the functional effect of H19 during cardiac differentiation. Expression levels of early cardiac-specific markers Nkx-2.5 and GATA4, cardiac contractile protein genes &alpha;-MHC and MLC-2v were determined by qRT-PCR and western lot. The levels of lncRNA H19 and miR-19b were detected by qRT-PCR. We further predicted the binding sequence of H19 and miR-19b by online softwares starBase v2.0 and TargetScan. The biological functions of H19 and Sox6 were evaluated by CCK-8 kit, cell cycle and apoptosis assay and caspase-3 activity.

Results: The expression levels of &alpha;-MHC, MLC-2v and H19 were upregulated, and miR-19b was downregulated significantly in mouse P19CL6 cells at the late stage of cardiac differentiation. Biological function analysis showed that knockdown of H19 promoted cell proliferation and inhibits cell apoptosis. H19 suppressed miR-19b expression and miR-19b targeted Sox6, which inhibited cell proliferation and promoted apoptosis in P19CL6 cells during late-stage cardiac differentiation. Importantly, Sox6 overexpression could reverse the positive effects of H19 knockdown on P19CL6 cells.

Conclusion: Downregulation of H19 promoted cell proliferation and inhibited cell apoptosis during late-stage cardiac differentiation by regulating the negative role of miR-19b in Sox6 expression, which suggested that the manipulation of H19 expression could serve as a potential strategy for heart disease.

No MeSH data available.


Related in: MedlinePlus