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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 multilineage differentiation capability of DPCs after transfection. After 21 d of multilineage differentiation induction, immunofluorescent staining indicated that DSPP (a1), LPL (a2), and collagen type II (a3) were extensively expressed in DPCs at P7 with XPC overexpression, strongly expressed in the nucleus and moderately expressed in the cytoplasm of DPCs. However, DSPP (a4), LPL (a5), and collagen type II (a6) revealed weak expression in the nucleus and cytoplasm of DPCs at P7 without transfection. Real-time PCR indicated that mRNA expression level of odontogenic markers (DMP1, DSPP), adipogenic markers (PPARγ2, LPL), and chondrogenic markers (collagen type II) increased significantly in DPCs at P3 and P7 with XPC overexpression compared with vector groups (b1–b5).
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fig5: Effect of XPC on the multilineage differentiation capability of DPCs after transfection. After 21 d of multilineage differentiation induction, immunofluorescent staining indicated that DSPP (a1), LPL (a2), and collagen type II (a3) were extensively expressed in DPCs at P7 with XPC overexpression, strongly expressed in the nucleus and moderately expressed in the cytoplasm of DPCs. However, DSPP (a4), LPL (a5), and collagen type II (a6) revealed weak expression in the nucleus and cytoplasm of DPCs at P7 without transfection. Real-time PCR indicated that mRNA expression level of odontogenic markers (DMP1, DSPP), adipogenic markers (PPARγ2, LPL), and chondrogenic markers (collagen type II) increased significantly in DPCs at P3 and P7 with XPC overexpression compared with vector groups (b1–b5).

Mentions: The odontogenic/adipogenic/chondrogenic differentiation capability of DPCs with XPC overexpression at P7 was investigated by immunofluorescent staining (Figures 5(a1)–5(a6)), and the mRNA expression of lineage related genes in DPCs at P3 and P7 was further confirmed by real-time PCR. After odontogenic/adipogenic/chondrogenic induction, immunofluorescent staining indicated that DSPP (Figure 5(a1)), LPL (Figure 5(a2)), and Collagen type II (Figure 5(a3)) were extensively expressed in DPCs at P7 with XPC overexpression, strongly expressed in the nucleus and moderately expressed in cytoplasm. However, the expression of DSPP (Figure 5(a4)), LPL (Figure 5(a5)), and Collagen type II (Figure 5(a6)) was barely detected in the DPCs at P7 without XPC transfection. Real-time PCR revealed that odontogenic markers (DMP1, DSPP), adipogenic markers (LPL, PPARγ2), and chondrogenic marker (Collagen type II) showed similar expression pattern, upregulated in DPCs with XPC overexpression at both P3 and P7 compared with the vector groups (Figures 5(b1)–5(b5), ∗p < 0.05, ∗∗p < 0.01). Therefore, XPC could effectively enhance the multilineage differentiation capability of DPCs with 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)

Effect of XPC on the multilineage differentiation capability of DPCs after transfection. After 21 d of multilineage differentiation induction, immunofluorescent staining indicated that DSPP (a1), LPL (a2), and collagen type II (a3) were extensively expressed in DPCs at P7 with XPC overexpression, strongly expressed in the nucleus and moderately expressed in the cytoplasm of DPCs. However, DSPP (a4), LPL (a5), and collagen type II (a6) revealed weak expression in the nucleus and cytoplasm of DPCs at P7 without transfection. Real-time PCR indicated that mRNA expression level of odontogenic markers (DMP1, DSPP), adipogenic markers (PPARγ2, LPL), and chondrogenic markers (collagen type II) increased significantly in DPCs at P3 and P7 with XPC overexpression compared with vector groups (b1–b5).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig5: Effect of XPC on the multilineage differentiation capability of DPCs after transfection. After 21 d of multilineage differentiation induction, immunofluorescent staining indicated that DSPP (a1), LPL (a2), and collagen type II (a3) were extensively expressed in DPCs at P7 with XPC overexpression, strongly expressed in the nucleus and moderately expressed in the cytoplasm of DPCs. However, DSPP (a4), LPL (a5), and collagen type II (a6) revealed weak expression in the nucleus and cytoplasm of DPCs at P7 without transfection. Real-time PCR indicated that mRNA expression level of odontogenic markers (DMP1, DSPP), adipogenic markers (PPARγ2, LPL), and chondrogenic markers (collagen type II) increased significantly in DPCs at P3 and P7 with XPC overexpression compared with vector groups (b1–b5).
Mentions: The odontogenic/adipogenic/chondrogenic differentiation capability of DPCs with XPC overexpression at P7 was investigated by immunofluorescent staining (Figures 5(a1)–5(a6)), and the mRNA expression of lineage related genes in DPCs at P3 and P7 was further confirmed by real-time PCR. After odontogenic/adipogenic/chondrogenic induction, immunofluorescent staining indicated that DSPP (Figure 5(a1)), LPL (Figure 5(a2)), and Collagen type II (Figure 5(a3)) were extensively expressed in DPCs at P7 with XPC overexpression, strongly expressed in the nucleus and moderately expressed in cytoplasm. However, the expression of DSPP (Figure 5(a4)), LPL (Figure 5(a5)), and Collagen type II (Figure 5(a6)) was barely detected in the DPCs at P7 without XPC transfection. Real-time PCR revealed that odontogenic markers (DMP1, DSPP), adipogenic markers (LPL, PPARγ2), and chondrogenic marker (Collagen type II) showed similar expression pattern, upregulated in DPCs with XPC overexpression at both P3 and P7 compared with the vector groups (Figures 5(b1)–5(b5), ∗p < 0.05, ∗∗p < 0.01). Therefore, XPC could effectively enhance the multilineage differentiation capability of DPCs with 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