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The PDGF-D/miR-106a/Twist1 pathway orchestrates epithelial-mesenchymal transition in gemcitabine resistance hepatoma cells.

Wang R, Li Y, Hou Y, Yang Q, Chen S, Wang X, Wang Z, Yang Y, Chen C, Wang Z, Wu Q - Oncotarget (2015)

Bottom Line: We found that PDGF-D is highly expressed in gemcitabine-resistant (GR) HCC cells.Notably, PDGF-D expression was associated with miR-106a and Twist1 in HCC patients.Therefore, inactivation of PDGF-D/Twist or activation of miR-106a could be a novel strategy for the treatment of HCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, China.

ABSTRACT
Emerging evidence demonstrates that platelet-derived growth factor-D (PDGF-D) plays a critical role in epithelial-mesenchymal transition (EMT) and drug resistance in hepatocellular carcinoma (HCC) cells. However, the underlying mechanism has not been fully elucidated. The objective is to explore the molecular mechanism of PDGF-D-mediated EMT in drug resistance HCC cells. To achieve our goal, we used multiple approaches including Western blotting, real-time RT-PCR, wound healing assay, invasion assay, luciferase activity assay, transfection, and immunohistochemistry. We found that PDGF-D is highly expressed in gemcitabine-resistant (GR) HCC cells. Moreover, PDGF-D markedly inhibited miR-106a expression and subsequently upregulated Twist1 expression. Notably, PDGF-D expression was associated with miR-106a and Twist1 in HCC patients. Our findings provide a possible molecular mechanism for understanding GR chemoresistance in HCC cells. Therefore, inactivation of PDGF-D/Twist or activation of miR-106a could be a novel strategy for the treatment of HCC.

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Related in: MedlinePlus

PDGF-D controls Twist1 expression(A) RT-PCR assay was performed to detect the mRNA level of Twist1 in indicated HCC cells. *p < 0.05, **p < 0.01 vs control. (B) Western blotting analysis was conducted to measure the expression of Twist1, PDGF-D in indicated HCC cells. (C) RT-PCR assay and Western blotting analysis were used to detect the expression of Twist1 in HepG2 and Huh-7 cells after PDGF-D cDNA transfection. *p < 0.05, **p < 0.01 vs control. (D) RT-PCR assay and Western blotting analysis were used to detect the expression of Twist1 in HepG2 GR and Huh-7 GR cells after PDGF-D siRNA transfection. *p < 0.05, **p < 0.01 vs control.
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Figure 5: PDGF-D controls Twist1 expression(A) RT-PCR assay was performed to detect the mRNA level of Twist1 in indicated HCC cells. *p < 0.05, **p < 0.01 vs control. (B) Western blotting analysis was conducted to measure the expression of Twist1, PDGF-D in indicated HCC cells. (C) RT-PCR assay and Western blotting analysis were used to detect the expression of Twist1 in HepG2 and Huh-7 cells after PDGF-D cDNA transfection. *p < 0.05, **p < 0.01 vs control. (D) RT-PCR assay and Western blotting analysis were used to detect the expression of Twist1 in HepG2 GR and Huh-7 GR cells after PDGF-D siRNA transfection. *p < 0.05, **p < 0.01 vs control.

Mentions: PDGF-D has been reported to be critically involved in GR-mediated EMT [11], we measured the expression of PDGF-D at mRNA and protein levels in HCC GR cells by RT-PCR and Western blotting, respectively. Consistent with our previous report [10], we observed a significantly increased PDGF-D at both mRNA and protein levels in HepG2 GR and Huh-7 GR cells (Figures 1C, 5B). It has been known that miR-106a plays a pivotal role in drug resistance [19]. Thus, we determine whether miR-106a has changes in HCC GR cells compared with the parental cells. Indeed, we observed the down-regulation of miR-106a in both HepG2 GR and Huh-7 GR cells (Figure 1C).


The PDGF-D/miR-106a/Twist1 pathway orchestrates epithelial-mesenchymal transition in gemcitabine resistance hepatoma cells.

Wang R, Li Y, Hou Y, Yang Q, Chen S, Wang X, Wang Z, Yang Y, Chen C, Wang Z, Wu Q - Oncotarget (2015)

PDGF-D controls Twist1 expression(A) RT-PCR assay was performed to detect the mRNA level of Twist1 in indicated HCC cells. *p < 0.05, **p < 0.01 vs control. (B) Western blotting analysis was conducted to measure the expression of Twist1, PDGF-D in indicated HCC cells. (C) RT-PCR assay and Western blotting analysis were used to detect the expression of Twist1 in HepG2 and Huh-7 cells after PDGF-D cDNA transfection. *p < 0.05, **p < 0.01 vs control. (D) RT-PCR assay and Western blotting analysis were used to detect the expression of Twist1 in HepG2 GR and Huh-7 GR cells after PDGF-D siRNA transfection. *p < 0.05, **p < 0.01 vs control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 5: PDGF-D controls Twist1 expression(A) RT-PCR assay was performed to detect the mRNA level of Twist1 in indicated HCC cells. *p < 0.05, **p < 0.01 vs control. (B) Western blotting analysis was conducted to measure the expression of Twist1, PDGF-D in indicated HCC cells. (C) RT-PCR assay and Western blotting analysis were used to detect the expression of Twist1 in HepG2 and Huh-7 cells after PDGF-D cDNA transfection. *p < 0.05, **p < 0.01 vs control. (D) RT-PCR assay and Western blotting analysis were used to detect the expression of Twist1 in HepG2 GR and Huh-7 GR cells after PDGF-D siRNA transfection. *p < 0.05, **p < 0.01 vs control.
Mentions: PDGF-D has been reported to be critically involved in GR-mediated EMT [11], we measured the expression of PDGF-D at mRNA and protein levels in HCC GR cells by RT-PCR and Western blotting, respectively. Consistent with our previous report [10], we observed a significantly increased PDGF-D at both mRNA and protein levels in HepG2 GR and Huh-7 GR cells (Figures 1C, 5B). It has been known that miR-106a plays a pivotal role in drug resistance [19]. Thus, we determine whether miR-106a has changes in HCC GR cells compared with the parental cells. Indeed, we observed the down-regulation of miR-106a in both HepG2 GR and Huh-7 GR cells (Figure 1C).

Bottom Line: We found that PDGF-D is highly expressed in gemcitabine-resistant (GR) HCC cells.Notably, PDGF-D expression was associated with miR-106a and Twist1 in HCC patients.Therefore, inactivation of PDGF-D/Twist or activation of miR-106a could be a novel strategy for the treatment of HCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, China.

ABSTRACT
Emerging evidence demonstrates that platelet-derived growth factor-D (PDGF-D) plays a critical role in epithelial-mesenchymal transition (EMT) and drug resistance in hepatocellular carcinoma (HCC) cells. However, the underlying mechanism has not been fully elucidated. The objective is to explore the molecular mechanism of PDGF-D-mediated EMT in drug resistance HCC cells. To achieve our goal, we used multiple approaches including Western blotting, real-time RT-PCR, wound healing assay, invasion assay, luciferase activity assay, transfection, and immunohistochemistry. We found that PDGF-D is highly expressed in gemcitabine-resistant (GR) HCC cells. Moreover, PDGF-D markedly inhibited miR-106a expression and subsequently upregulated Twist1 expression. Notably, PDGF-D expression was associated with miR-106a and Twist1 in HCC patients. Our findings provide a possible molecular mechanism for understanding GR chemoresistance in HCC cells. Therefore, inactivation of PDGF-D/Twist or activation of miR-106a could be a novel strategy for the treatment of HCC.

Show MeSH
Related in: MedlinePlus