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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

Effect of XPC on the expression of Oct-4, Sox2, and c-Myc in DPCs at various passages. Immunofluorescent staining revealed that XPC (a1), Oct-4 (a2), Sox2 (a3), and c-Myc (a4) were strongly expressed in DPCs with XPC overexpression at P7, mainly located in nucleus and moderately expressed in cytoplasm, whereas XPC (a5), Oct-4 (a6), Sox2 (a7), and c-Myc (a8) were barely detected in DPCs at P7 without transfection. Real-time PCR indicated that the mRNA expression of XPC, Oct-4, Sox2, and c-Myc was upregulated significantly in DPCs with XPC overexpression at both P3 and P7 compared with vector groups (∗∗p < 0.001). There is no significant difference between nontransfected group and vector group (p > 0.05) (b1–b3).
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fig4: Effect of XPC on the expression of Oct-4, Sox2, and c-Myc in DPCs at various passages. Immunofluorescent staining revealed that XPC (a1), Oct-4 (a2), Sox2 (a3), and c-Myc (a4) were strongly expressed in DPCs with XPC overexpression at P7, mainly located in nucleus and moderately expressed in cytoplasm, whereas XPC (a5), Oct-4 (a6), Sox2 (a7), and c-Myc (a8) were barely detected in DPCs at P7 without transfection. Real-time PCR indicated that the mRNA expression of XPC, Oct-4, Sox2, and c-Myc was upregulated significantly in DPCs with XPC overexpression at both P3 and P7 compared with vector groups (∗∗p < 0.001). There is no significant difference between nontransfected group and vector group (p > 0.05) (b1–b3).

Mentions: Real-time PCR indicated that the mRNA level of Oct-4, Sox2, and c-Myc was significantly upregulated in DPCs with XPC transfection at both P3 and P7 (Figures 4(b1)–4(b3), ∗∗p < 0.001). Similarly, immunofluorescent staining revealed that XPC (Figure 4(a1)), Oct-4 (Figure 4(a2)), Sox2 (Figure 4(a3)), and c-Myc (Figure 4(a4)) were strongly expressed in DPCs with XPC overexpression at P7, mainly located in nucleus and moderately expressed in cytoplasm. Whereas XPC (Figure 4(a5)), Oct-4 (Figure 4(a6)), Sox2 (Figure 4(a7)), and c-Myc (Figure 4(a8)) were barely detected in DPCs at P7 without transfection. Oct-4, Sox2, and c-Myc showed extensive expression in DPCs at P3 with/without XPC expression (data not shown).


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 the expression of Oct-4, Sox2, and c-Myc in DPCs at various passages. Immunofluorescent staining revealed that XPC (a1), Oct-4 (a2), Sox2 (a3), and c-Myc (a4) were strongly expressed in DPCs with XPC overexpression at P7, mainly located in nucleus and moderately expressed in cytoplasm, whereas XPC (a5), Oct-4 (a6), Sox2 (a7), and c-Myc (a8) were barely detected in DPCs at P7 without transfection. Real-time PCR indicated that the mRNA expression of XPC, Oct-4, Sox2, and c-Myc was upregulated significantly in DPCs with XPC overexpression at both P3 and P7 compared with vector groups (∗∗p < 0.001). There is no significant difference between nontransfected group and vector group (p > 0.05) (b1–b3).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Effect of XPC on the expression of Oct-4, Sox2, and c-Myc in DPCs at various passages. Immunofluorescent staining revealed that XPC (a1), Oct-4 (a2), Sox2 (a3), and c-Myc (a4) were strongly expressed in DPCs with XPC overexpression at P7, mainly located in nucleus and moderately expressed in cytoplasm, whereas XPC (a5), Oct-4 (a6), Sox2 (a7), and c-Myc (a8) were barely detected in DPCs at P7 without transfection. Real-time PCR indicated that the mRNA expression of XPC, Oct-4, Sox2, and c-Myc was upregulated significantly in DPCs with XPC overexpression at both P3 and P7 compared with vector groups (∗∗p < 0.001). There is no significant difference between nontransfected group and vector group (p > 0.05) (b1–b3).
Mentions: Real-time PCR indicated that the mRNA level of Oct-4, Sox2, and c-Myc was significantly upregulated in DPCs with XPC transfection at both P3 and P7 (Figures 4(b1)–4(b3), ∗∗p < 0.001). Similarly, immunofluorescent staining revealed that XPC (Figure 4(a1)), Oct-4 (Figure 4(a2)), Sox2 (Figure 4(a3)), and c-Myc (Figure 4(a4)) were strongly expressed in DPCs with XPC overexpression at P7, mainly located in nucleus and moderately expressed in cytoplasm. Whereas XPC (Figure 4(a5)), Oct-4 (Figure 4(a6)), Sox2 (Figure 4(a7)), and c-Myc (Figure 4(a8)) were barely detected in DPCs at P7 without transfection. Oct-4, Sox2, and c-Myc showed extensive expression in DPCs at P3 with/without XPC expression (data not shown).

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