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Serum Dickkopf-1 as a Biomarker for the Diagnosis of Hepatocellular Carcinoma.

Kim SU, Park JH, Kim HS, Lee JM, Lee HG, Kim H, Choi SH, Baek S, Kim BK, Park JY, Kim do Y, Ahn SH, Lee JD, Han KH - Yonsei Med. J. (2015)

Bottom Line: When serum DKK-1 levels were assessed in 370 participants (217 with HCC and 153 without), it was significantly higher in HCC patients than in control groups (median 1.48 ng/mL vs. 0.90 ng/mL, p<0.001).When three biomarkers were combined (DKK-1 plus AFP plus DCP), they showed significantly higher AUC (AUC=0.952) than single marker, DKK-1 plus AFP, or DKK-1 plus DCP (all p<0.001).Serum DKK-1 could complement the diagnostic accuracy of AFP and DCP.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT

Purpose: Dickkopf-1 (DKK-1) is a Wnt/β-catenin signaling pathway inhibitor. We investigated whether DKK-1 is related to progression in hepatocellular carcinoma (HCC) cells and HCC patients.

Materials and methods: In vitro reverse-transcription polymerase chain reaction (RT-PCR), wound healing assays, invasion assays, and ELISAs of patient serum samples were employed. The diagnostic accuracy of the serum DKK-1 ELISA was assessed using receiver operating characteristic (ROC) curves and area under ROC (AUC) analyses.

Results: RT-PCR showed high DKK-1 expression in Hep3B and low in 293 cells. Similarly, the secreted DKK-1 concentration in the culture media was high in Hep3B and low in 293 cells. Wound healing and invasion assays using 293, Huh7, and Hep3B cells showed that DKK-1 overexpression promoted cell migration and invasion, whereas DKK-1 knock-down inhibited them. When serum DKK-1 levels were assessed in 370 participants (217 with HCC and 153 without), it was significantly higher in HCC patients than in control groups (median 1.48 ng/mL vs. 0.90 ng/mL, p<0.001). The optimum DKK-1 cutoff level was 1.01 ng/mL (AUC=0.829; sensitivity 90.7%; specificity 62.0%). Although DKK-1 had a higher AUC than alpha-fetoprotein (AFP) and des-gamma-carboxy prothrombin (DCP) (AUC=0.829 vs. 0.794 and 0.815, respectively), they were statistically similar (all p>0.05). When three biomarkers were combined (DKK-1 plus AFP plus DCP), they showed significantly higher AUC (AUC=0.952) than single marker, DKK-1 plus AFP, or DKK-1 plus DCP (all p<0.001).

Conclusion: DKK-1 might be a key regulator in HCC progression and a potential therapeutic target in HCC. Serum DKK-1 could complement the diagnostic accuracy of AFP and DCP.

No MeSH data available.


Related in: MedlinePlus

DKK-1 expression in Hep3B, Huh7, and 293 cells. (A) RT-PCR analysis showed high DKK-1 mRNA expression in Hep3B cells, low in 293 cells, and none in the negative control. (B) The DKK-1 concentration secreted in the culture media, determined by ELISA, was high in Hep3B cells, moderate in Huh7 cells, and low in 293 cells. (C) Following DKK-1 transfection, luciferase assays revealed that the β-catenin transcriptional activity was induced in 293 cells. *p<0.01. DKK-1, dickkopf-1.
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Figure 1: DKK-1 expression in Hep3B, Huh7, and 293 cells. (A) RT-PCR analysis showed high DKK-1 mRNA expression in Hep3B cells, low in 293 cells, and none in the negative control. (B) The DKK-1 concentration secreted in the culture media, determined by ELISA, was high in Hep3B cells, moderate in Huh7 cells, and low in 293 cells. (C) Following DKK-1 transfection, luciferase assays revealed that the β-catenin transcriptional activity was induced in 293 cells. *p<0.01. DKK-1, dickkopf-1.

Mentions: To determine the DKK-1 expression in HCC cells, reverse transcription-PCR (RT-PCR) and ELISA were performed in Hep3B and Huh7 cells with 293 cells as a negative control. RT-PCR analysis showed high DKK-1 mRNA expression in Hep3B cells and low in 293 cells (Fig. 1A). Consistent with these findings, the secreted DKK-1 concentration in the culture media was high in Hep3B cells, moderate in Huh7 cells, and low in 293 cells (Fig. 1B). Luciferase assays revealed that β-catenin transcriptional activity increased significantly in 293 cells (approximately 2.5-fold) following DKK-1 overexpression. In Hep3B cells, however, the activity remained comparable between mock and DKK-1 overexpression (Fig. 1C).


Serum Dickkopf-1 as a Biomarker for the Diagnosis of Hepatocellular Carcinoma.

Kim SU, Park JH, Kim HS, Lee JM, Lee HG, Kim H, Choi SH, Baek S, Kim BK, Park JY, Kim do Y, Ahn SH, Lee JD, Han KH - Yonsei Med. J. (2015)

DKK-1 expression in Hep3B, Huh7, and 293 cells. (A) RT-PCR analysis showed high DKK-1 mRNA expression in Hep3B cells, low in 293 cells, and none in the negative control. (B) The DKK-1 concentration secreted in the culture media, determined by ELISA, was high in Hep3B cells, moderate in Huh7 cells, and low in 293 cells. (C) Following DKK-1 transfection, luciferase assays revealed that the β-catenin transcriptional activity was induced in 293 cells. *p<0.01. DKK-1, dickkopf-1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4541659&req=5

Figure 1: DKK-1 expression in Hep3B, Huh7, and 293 cells. (A) RT-PCR analysis showed high DKK-1 mRNA expression in Hep3B cells, low in 293 cells, and none in the negative control. (B) The DKK-1 concentration secreted in the culture media, determined by ELISA, was high in Hep3B cells, moderate in Huh7 cells, and low in 293 cells. (C) Following DKK-1 transfection, luciferase assays revealed that the β-catenin transcriptional activity was induced in 293 cells. *p<0.01. DKK-1, dickkopf-1.
Mentions: To determine the DKK-1 expression in HCC cells, reverse transcription-PCR (RT-PCR) and ELISA were performed in Hep3B and Huh7 cells with 293 cells as a negative control. RT-PCR analysis showed high DKK-1 mRNA expression in Hep3B cells and low in 293 cells (Fig. 1A). Consistent with these findings, the secreted DKK-1 concentration in the culture media was high in Hep3B cells, moderate in Huh7 cells, and low in 293 cells (Fig. 1B). Luciferase assays revealed that β-catenin transcriptional activity increased significantly in 293 cells (approximately 2.5-fold) following DKK-1 overexpression. In Hep3B cells, however, the activity remained comparable between mock and DKK-1 overexpression (Fig. 1C).

Bottom Line: When serum DKK-1 levels were assessed in 370 participants (217 with HCC and 153 without), it was significantly higher in HCC patients than in control groups (median 1.48 ng/mL vs. 0.90 ng/mL, p<0.001).When three biomarkers were combined (DKK-1 plus AFP plus DCP), they showed significantly higher AUC (AUC=0.952) than single marker, DKK-1 plus AFP, or DKK-1 plus DCP (all p<0.001).Serum DKK-1 could complement the diagnostic accuracy of AFP and DCP.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT

Purpose: Dickkopf-1 (DKK-1) is a Wnt/β-catenin signaling pathway inhibitor. We investigated whether DKK-1 is related to progression in hepatocellular carcinoma (HCC) cells and HCC patients.

Materials and methods: In vitro reverse-transcription polymerase chain reaction (RT-PCR), wound healing assays, invasion assays, and ELISAs of patient serum samples were employed. The diagnostic accuracy of the serum DKK-1 ELISA was assessed using receiver operating characteristic (ROC) curves and area under ROC (AUC) analyses.

Results: RT-PCR showed high DKK-1 expression in Hep3B and low in 293 cells. Similarly, the secreted DKK-1 concentration in the culture media was high in Hep3B and low in 293 cells. Wound healing and invasion assays using 293, Huh7, and Hep3B cells showed that DKK-1 overexpression promoted cell migration and invasion, whereas DKK-1 knock-down inhibited them. When serum DKK-1 levels were assessed in 370 participants (217 with HCC and 153 without), it was significantly higher in HCC patients than in control groups (median 1.48 ng/mL vs. 0.90 ng/mL, p<0.001). The optimum DKK-1 cutoff level was 1.01 ng/mL (AUC=0.829; sensitivity 90.7%; specificity 62.0%). Although DKK-1 had a higher AUC than alpha-fetoprotein (AFP) and des-gamma-carboxy prothrombin (DCP) (AUC=0.829 vs. 0.794 and 0.815, respectively), they were statistically similar (all p>0.05). When three biomarkers were combined (DKK-1 plus AFP plus DCP), they showed significantly higher AUC (AUC=0.952) than single marker, DKK-1 plus AFP, or DKK-1 plus DCP (all p<0.001).

Conclusion: DKK-1 might be a key regulator in HCC progression and a potential therapeutic target in HCC. Serum DKK-1 could complement the diagnostic accuracy of AFP and DCP.

No MeSH data available.


Related in: MedlinePlus