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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.


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Effect of XPC on cell proliferation and apoptosis and cell cycle and senescence of DPCs with in vitro culture. CCK8 revealed that cell proliferation of transfected DPCs at both P3 and P7 was significantly enhanced (b1, ∗∗p < 0.001). The result of FACS (a1–a6) demonstrated that the cell number of transfected DPCs at P3 and P7 in G0/G1 was significantly downregulated compared with the vector groups; DPCs were arrested in G2/M and S phase. The percentage of PI = (S + G2/M) value (b3, ∗p < 0.05) and telomerase activity (b4, ∗∗p < 0.001) of transfected DPCs at both P3 and P7 was significantly upregulated, albeit the apoptosis rate was significantly downregulated (b2, ∗p < 0.05).
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fig3: Effect of XPC on cell proliferation and apoptosis and cell cycle and senescence of DPCs with in vitro culture. CCK8 revealed that cell proliferation of transfected DPCs at both P3 and P7 was significantly enhanced (b1, ∗∗p < 0.001). The result of FACS (a1–a6) demonstrated that the cell number of transfected DPCs at P3 and P7 in G0/G1 was significantly downregulated compared with the vector groups; DPCs were arrested in G2/M and S phase. The percentage of PI = (S + G2/M) value (b3, ∗p < 0.05) and telomerase activity (b4, ∗∗p < 0.001) of transfected DPCs at both P3 and P7 was significantly upregulated, albeit the apoptosis rate was significantly downregulated (b2, ∗p < 0.05).

Mentions: To investigate the biological function of XPC in DPCs, CCK8 and FACS were applied to DPCs with XPC overexpression. CCK8 revealed that cell proliferation of transfected DPCs at both P3 and P7 was significantly enhanced (Figure 3(b1), ∗∗p < 0.001), whereas there is no significant difference between control group and vector group (p > 0.05). The result of FACS demonstrated that the cell number of transfected DPCs at P3 and P7 in G0/G1 was significantly downregulated compared with the vector groups; DPCs were arrested in G2/M and S phase. The percentage of PI = (S + G2/M) value and telomerase activity of transfected DPCs at both P3 and P7 was significantly upregulated, albeit the apoptosis rate was significantly downregulated (Figures 3(b2)–3(b4), Table 3, p < 0.05). Therefore, XPC could enhance cell proliferation and telomerase activity of DPCs, arrested cells in G2/M and S phase, and upregulated PI value, albeit suppressed apoptosis, suggesting the role XPC played in the maintenance of pluripotency in DPCs.


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)

Effect of XPC on cell proliferation and apoptosis and cell cycle and senescence of DPCs with in vitro culture. CCK8 revealed that cell proliferation of transfected DPCs at both P3 and P7 was significantly enhanced (b1, ∗∗p < 0.001). The result of FACS (a1–a6) demonstrated that the cell number of transfected DPCs at P3 and P7 in G0/G1 was significantly downregulated compared with the vector groups; DPCs were arrested in G2/M and S phase. The percentage of PI = (S + G2/M) value (b3, ∗p < 0.05) and telomerase activity (b4, ∗∗p < 0.001) of transfected DPCs at both P3 and P7 was significantly upregulated, albeit the apoptosis rate was significantly downregulated (b2, ∗p < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4834411&req=5

fig3: Effect of XPC on cell proliferation and apoptosis and cell cycle and senescence of DPCs with in vitro culture. CCK8 revealed that cell proliferation of transfected DPCs at both P3 and P7 was significantly enhanced (b1, ∗∗p < 0.001). The result of FACS (a1–a6) demonstrated that the cell number of transfected DPCs at P3 and P7 in G0/G1 was significantly downregulated compared with the vector groups; DPCs were arrested in G2/M and S phase. The percentage of PI = (S + G2/M) value (b3, ∗p < 0.05) and telomerase activity (b4, ∗∗p < 0.001) of transfected DPCs at both P3 and P7 was significantly upregulated, albeit the apoptosis rate was significantly downregulated (b2, ∗p < 0.05).
Mentions: To investigate the biological function of XPC in DPCs, CCK8 and FACS were applied to DPCs with XPC overexpression. CCK8 revealed that cell proliferation of transfected DPCs at both P3 and P7 was significantly enhanced (Figure 3(b1), ∗∗p < 0.001), whereas there is no significant difference between control group and vector group (p > 0.05). The result of FACS demonstrated that the cell number of transfected DPCs at P3 and P7 in G0/G1 was significantly downregulated compared with the vector groups; DPCs were arrested in G2/M and S phase. The percentage of PI = (S + G2/M) value and telomerase activity of transfected DPCs at both P3 and P7 was significantly upregulated, albeit the apoptosis rate was significantly downregulated (Figures 3(b2)–3(b4), Table 3, p < 0.05). Therefore, XPC could enhance cell proliferation and telomerase activity of DPCs, arrested cells in G2/M and S phase, and upregulated PI value, albeit suppressed apoptosis, suggesting the role XPC played in the maintenance of pluripotency in DPCs.

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