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XPC Promotes Pluripotency of Human Dental Pulp Cells through Regulation of Oct-4/Sox2/c-Myc.

Liu L, Peng Z, Xu Z, Wei X - Stem Cells Int (2016)

Bottom Line: XPC, Oct-4, Sox2, and c-Myc were downregulated at P7 compared with P3 in DPCs with long-term culture.Oct-4/Sox2/c-Myc were significantly upregulated, and multilineage differentiation in DPCs with XPC overexpression was enhanced after transfection.Conclusions.

View Article: PubMed Central - PubMed

Affiliation: Operative Dentistry and Endodontics, Guanghua School of Stomatology, Affiliated Stomatological Hospital, Guangdong Province Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong 510055, China.

ABSTRACT
Introduction. Xeroderma pigmentosum group C (XPC), essential component of multisubunit stem cell coactivator complex (SCC), functions as the critical factor modulating pluripotency and genome integrity through interaction with Oct-4/Sox2. However, its specific role in regulating pluripotency and multilineage differentiation of human dental pulp cells (DPCs) remains unknown. Methods. To elucidate the functional role XPC played in pluripotency and multilineage differentiation of DPCs, expressions of XPC in DPCs with long-term culture were examined by real-time PCR and western blot. DPCs were transfected with lentiviral-mediated human XPC gene; then transfection rate was investigated by real-time PCR and western blot. Cell cycle, apoptosis, proliferation, senescence, multilineage differentiation, and expression of Oct-4/Sox2/c-Myc in transfected DPCs were examined. Results. XPC, Oct-4, Sox2, and c-Myc were downregulated at P7 compared with P3 in DPCs with long-term culture. XPC genes were upregulated in DPCs at P2 after transfection and maintained high expression level at P3 and P7. Cell proliferation, PI value, and telomerase activity were enhanced, whereas apoptosis was suppressed in transfected DPCs. Oct-4/Sox2/c-Myc were significantly upregulated, and multilineage differentiation in DPCs with XPC overexpression was enhanced after transfection. Conclusions. XPC plays an essential role in the modulation of pluripotency and multilineage differentiation of DPCs through regulation of Oct-4/Sox2/c-Myc.

No MeSH data available.


Related in: MedlinePlus

Establishment of XPC overexpression DPCs model. Expression of pCDH-CMV-XPC-EF1-copGFP plasmid in HEK293T cells and DPCs at P2 (a1–a6). Strong green fluorescence was detected in HEK293T cells (a2, ×50) and DPCs (a5, ×100) after transfection. Combined figures of the bright field and fluorescence figure (a3, a6). Western blot and real-time PCR showed that the expression of XPC was enhanced in XPC+/DPCs at P2 at both protein and mRNA level compared with vector group, and XPC maintained high expression level in DPCs at P3 and P7 (∗∗p < 0.01) (b1, b2).
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fig2: Establishment of XPC overexpression DPCs model. Expression of pCDH-CMV-XPC-EF1-copGFP plasmid in HEK293T cells and DPCs at P2 (a1–a6). Strong green fluorescence was detected in HEK293T cells (a2, ×50) and DPCs (a5, ×100) after transfection. Combined figures of the bright field and fluorescence figure (a3, a6). Western blot and real-time PCR showed that the expression of XPC was enhanced in XPC+/DPCs at P2 at both protein and mRNA level compared with vector group, and XPC maintained high expression level in DPCs at P3 and P7 (∗∗p < 0.01) (b1, b2).

Mentions: Expression of pCDH-CMV-XPC-EF1-copGFP plasmid in HEK293T cells and DPCs was shown in Figure 2. Strong green fluorescence could be viewed in HEK293T cells after transfection (Figure 2(a2)). All DPCs with XPC overexpression showed green fluorescent staining (Figure 2(a5)). The result of real-time PCR revealed that XPC mRNA was significantly higher in XPC transfected DPCs at P2, P3, and P7 compared with the vector group (Figure 2(b2), ∗∗p < 0.001), whereas there is no significant difference between nontransfected and vector groups (p > 0.05). Western blot (Figure 2(b1)) demonstrated that the protein expression of XPC was strongly upregulated in DPCs at P2, P3, and P7 after transfection, which agreed with the result of real-time PCR (Figure 2(b2)). These results indicated that the XPC overexpression DPCs model was successfully established, which could be passaged and XPC maintained consistently high expression level in transfected DPCs even after long-term in vitro culture.


XPC Promotes Pluripotency of Human Dental Pulp Cells through Regulation of Oct-4/Sox2/c-Myc.

Liu L, Peng Z, Xu Z, Wei X - Stem Cells Int (2016)

Establishment of XPC overexpression DPCs model. Expression of pCDH-CMV-XPC-EF1-copGFP plasmid in HEK293T cells and DPCs at P2 (a1–a6). Strong green fluorescence was detected in HEK293T cells (a2, ×50) and DPCs (a5, ×100) after transfection. Combined figures of the bright field and fluorescence figure (a3, a6). Western blot and real-time PCR showed that the expression of XPC was enhanced in XPC+/DPCs at P2 at both protein and mRNA level compared with vector group, and XPC maintained high expression level in DPCs at P3 and P7 (∗∗p < 0.01) (b1, b2).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig2: Establishment of XPC overexpression DPCs model. Expression of pCDH-CMV-XPC-EF1-copGFP plasmid in HEK293T cells and DPCs at P2 (a1–a6). Strong green fluorescence was detected in HEK293T cells (a2, ×50) and DPCs (a5, ×100) after transfection. Combined figures of the bright field and fluorescence figure (a3, a6). Western blot and real-time PCR showed that the expression of XPC was enhanced in XPC+/DPCs at P2 at both protein and mRNA level compared with vector group, and XPC maintained high expression level in DPCs at P3 and P7 (∗∗p < 0.01) (b1, b2).
Mentions: Expression of pCDH-CMV-XPC-EF1-copGFP plasmid in HEK293T cells and DPCs was shown in Figure 2. Strong green fluorescence could be viewed in HEK293T cells after transfection (Figure 2(a2)). All DPCs with XPC overexpression showed green fluorescent staining (Figure 2(a5)). The result of real-time PCR revealed that XPC mRNA was significantly higher in XPC transfected DPCs at P2, P3, and P7 compared with the vector group (Figure 2(b2), ∗∗p < 0.001), whereas there is no significant difference between nontransfected and vector groups (p > 0.05). Western blot (Figure 2(b1)) demonstrated that the protein expression of XPC was strongly upregulated in DPCs at P2, P3, and P7 after transfection, which agreed with the result of real-time PCR (Figure 2(b2)). These results indicated that the XPC overexpression DPCs model was successfully established, which could be passaged and XPC maintained consistently high expression level in transfected DPCs even after long-term in vitro culture.

Bottom Line: XPC, Oct-4, Sox2, and c-Myc were downregulated at P7 compared with P3 in DPCs with long-term culture.Oct-4/Sox2/c-Myc were significantly upregulated, and multilineage differentiation in DPCs with XPC overexpression was enhanced after transfection.Conclusions.

View Article: PubMed Central - PubMed

Affiliation: Operative Dentistry and Endodontics, Guanghua School of Stomatology, Affiliated Stomatological Hospital, Guangdong Province Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong 510055, China.

ABSTRACT
Introduction. Xeroderma pigmentosum group C (XPC), essential component of multisubunit stem cell coactivator complex (SCC), functions as the critical factor modulating pluripotency and genome integrity through interaction with Oct-4/Sox2. However, its specific role in regulating pluripotency and multilineage differentiation of human dental pulp cells (DPCs) remains unknown. Methods. To elucidate the functional role XPC played in pluripotency and multilineage differentiation of DPCs, expressions of XPC in DPCs with long-term culture were examined by real-time PCR and western blot. DPCs were transfected with lentiviral-mediated human XPC gene; then transfection rate was investigated by real-time PCR and western blot. Cell cycle, apoptosis, proliferation, senescence, multilineage differentiation, and expression of Oct-4/Sox2/c-Myc in transfected DPCs were examined. Results. XPC, Oct-4, Sox2, and c-Myc were downregulated at P7 compared with P3 in DPCs with long-term culture. XPC genes were upregulated in DPCs at P2 after transfection and maintained high expression level at P3 and P7. Cell proliferation, PI value, and telomerase activity were enhanced, whereas apoptosis was suppressed in transfected DPCs. Oct-4/Sox2/c-Myc were significantly upregulated, and multilineage differentiation in DPCs with XPC overexpression was enhanced after transfection. Conclusions. XPC plays an essential role in the modulation of pluripotency and multilineage differentiation of DPCs through regulation of Oct-4/Sox2/c-Myc.

No MeSH data available.


Related in: MedlinePlus