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The significance of Notch1 compared with Notch3 in high metastasis and poor overall survival in hepatocellular carcinoma.

Zhou L, Zhang N, Song W, You N, Li Q, Sun W, Zhang Y, Wang D, Dou K - PLoS ONE (2013)

Bottom Line: Kaplan-Meier curves demonstrated that patients with high Notch1 or Notch3 expression were at a significantly increased risk for shortened survival time.In vitro, the down-regulation of Notch1 decreased the migration and invasion capacities of HCC cells by regulating CD44v6, E-cadherin, MMP-2, MMP-9, and uPA via the COX-2 and ERK1/2 pathways.Down-regulation of Notch3 only decreased the invasion capacity of HCC cells by regulating MMP-2 and MMP-9 via the ERK1/2 pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shannxi, People's Republic of China.

ABSTRACT

Background: The prognosis for patients with hepatocellular carcinoma (HCC) is poor, and the mechanisms underlying the development of HCC remain unclear. Notch1 and Notch3 may be involved in malignant transformation, although their roles remain unknown.

Materials and methods: HCC tissues were stained with anti-Notch1 or -Notch3 antibody. The migration and invasion capacities of the cells were measured with transwell cell culture chambers. RT-PCR was used to measure the expression of Notch1 and Notch3 mRNA. Additionally, western blot analysis was used to assess the protein expression of Notch1, Notch3, CD44v6, E-cadherin, matrix metalloproteinase-2 (MMP-2), MMP-9, and urokinase-type plasminogen activator (uPA). RNA interference was used to down-regulate the expression of Notch1 and Notch3. Cell viability was assessed using MTT.

Results: Based on immunohistochemistry, high Notch1 expression was correlated with tumor size, tumor grade, metastasis, venous invasion and AJCC TNM stage. High Notch3 expression was only strongly correlated with metastasis, venous invasion and satellite lesions. Kaplan-Meier curves demonstrated that patients with high Notch1 or Notch3 expression were at a significantly increased risk for shortened survival time. In vitro, the down-regulation of Notch1 decreased the migration and invasion capacities of HCC cells by regulating CD44v6, E-cadherin, MMP-2, MMP-9, and uPA via the COX-2 and ERK1/2 pathways. Down-regulation of Notch3 only decreased the invasion capacity of HCC cells by regulating MMP-2 and MMP-9 via the ERK1/2 pathway.

Conclusions: Based on the migration and invasion of HCC, we hypothesize that targeting Notch1 may be more useful than Notch3 for designing novel preventive and therapeutic strategies for HCC in the near future.

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Related in: MedlinePlus

Down-regulation of Notch1 or Notch3 can decrease the migration and invasion of HepG2 and MHCC97H cells in vitro.(a–c) Migrated HCC cells analyzed by transwell assays compared with siRNA controls. (d–f) Invaded HCC cells analyzed by transwell assays compared with siRNA controls. The data are presented as the mean ± SD, *P<0.05 compared with control siRNA-transfected HepG2 cells; #P<0.05 compared with control siRNA-transfected MHCC97H cells. NT: No transfection; Cs: control siRNA transfection; N1s: Notch1 siRNA transfection; N3s: Notch3 siRNA transfection.
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pone-0057382-g002: Down-regulation of Notch1 or Notch3 can decrease the migration and invasion of HepG2 and MHCC97H cells in vitro.(a–c) Migrated HCC cells analyzed by transwell assays compared with siRNA controls. (d–f) Invaded HCC cells analyzed by transwell assays compared with siRNA controls. The data are presented as the mean ± SD, *P<0.05 compared with control siRNA-transfected HepG2 cells; #P<0.05 compared with control siRNA-transfected MHCC97H cells. NT: No transfection; Cs: control siRNA transfection; N1s: Notch1 siRNA transfection; N3s: Notch3 siRNA transfection.

Mentions: Using transwell cell culture chambers, we measured the migration and invasion of Notch1 and Notch3 siRNA-transfected HepG2 and MHCC97H cells. HepG2 and MHCC97H cells transfected with control siRNA were used as controls. Only Notch1 down-regulation significantly decreased the numbers of migratory HCC cells (Fig. 2a–c). However, Notch1 or Notch3 down-regulation significantly decreased the numbers of invasive HCC cells (Fig. 2d–f). These data indicated that Notch1 down-regulation can reduce the migration and invasion of HCC cells. However, Notch3 down-regulation had no effect on migration and could only reduce invasion in HCC cells. Furthermore, to determine the potential mechanisms of Notch1 and Notch3 in the migration and invasion of HCC cells, we examined the effect of down-regulated Notch1 and Notch3 on metastasis-associated molecules, such as CD44v6, E-cadherin, MMP-2, MMP-9, and uPA. As Fig. 3a shows, Notch1 down-regulation decreases the protein expression of CD44v6, MMP-2, MMP-9, and uPA while increasing the protein expression of E-cadherin in HCC cells. However, Notch3 down-regulation only decreased the protein expression of MMP-2, MMP-9, and uPA.


The significance of Notch1 compared with Notch3 in high metastasis and poor overall survival in hepatocellular carcinoma.

Zhou L, Zhang N, Song W, You N, Li Q, Sun W, Zhang Y, Wang D, Dou K - PLoS ONE (2013)

Down-regulation of Notch1 or Notch3 can decrease the migration and invasion of HepG2 and MHCC97H cells in vitro.(a–c) Migrated HCC cells analyzed by transwell assays compared with siRNA controls. (d–f) Invaded HCC cells analyzed by transwell assays compared with siRNA controls. The data are presented as the mean ± SD, *P<0.05 compared with control siRNA-transfected HepG2 cells; #P<0.05 compared with control siRNA-transfected MHCC97H cells. NT: No transfection; Cs: control siRNA transfection; N1s: Notch1 siRNA transfection; N3s: Notch3 siRNA transfection.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3585338&req=5

pone-0057382-g002: Down-regulation of Notch1 or Notch3 can decrease the migration and invasion of HepG2 and MHCC97H cells in vitro.(a–c) Migrated HCC cells analyzed by transwell assays compared with siRNA controls. (d–f) Invaded HCC cells analyzed by transwell assays compared with siRNA controls. The data are presented as the mean ± SD, *P<0.05 compared with control siRNA-transfected HepG2 cells; #P<0.05 compared with control siRNA-transfected MHCC97H cells. NT: No transfection; Cs: control siRNA transfection; N1s: Notch1 siRNA transfection; N3s: Notch3 siRNA transfection.
Mentions: Using transwell cell culture chambers, we measured the migration and invasion of Notch1 and Notch3 siRNA-transfected HepG2 and MHCC97H cells. HepG2 and MHCC97H cells transfected with control siRNA were used as controls. Only Notch1 down-regulation significantly decreased the numbers of migratory HCC cells (Fig. 2a–c). However, Notch1 or Notch3 down-regulation significantly decreased the numbers of invasive HCC cells (Fig. 2d–f). These data indicated that Notch1 down-regulation can reduce the migration and invasion of HCC cells. However, Notch3 down-regulation had no effect on migration and could only reduce invasion in HCC cells. Furthermore, to determine the potential mechanisms of Notch1 and Notch3 in the migration and invasion of HCC cells, we examined the effect of down-regulated Notch1 and Notch3 on metastasis-associated molecules, such as CD44v6, E-cadherin, MMP-2, MMP-9, and uPA. As Fig. 3a shows, Notch1 down-regulation decreases the protein expression of CD44v6, MMP-2, MMP-9, and uPA while increasing the protein expression of E-cadherin in HCC cells. However, Notch3 down-regulation only decreased the protein expression of MMP-2, MMP-9, and uPA.

Bottom Line: Kaplan-Meier curves demonstrated that patients with high Notch1 or Notch3 expression were at a significantly increased risk for shortened survival time.In vitro, the down-regulation of Notch1 decreased the migration and invasion capacities of HCC cells by regulating CD44v6, E-cadherin, MMP-2, MMP-9, and uPA via the COX-2 and ERK1/2 pathways.Down-regulation of Notch3 only decreased the invasion capacity of HCC cells by regulating MMP-2 and MMP-9 via the ERK1/2 pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shannxi, People's Republic of China.

ABSTRACT

Background: The prognosis for patients with hepatocellular carcinoma (HCC) is poor, and the mechanisms underlying the development of HCC remain unclear. Notch1 and Notch3 may be involved in malignant transformation, although their roles remain unknown.

Materials and methods: HCC tissues were stained with anti-Notch1 or -Notch3 antibody. The migration and invasion capacities of the cells were measured with transwell cell culture chambers. RT-PCR was used to measure the expression of Notch1 and Notch3 mRNA. Additionally, western blot analysis was used to assess the protein expression of Notch1, Notch3, CD44v6, E-cadherin, matrix metalloproteinase-2 (MMP-2), MMP-9, and urokinase-type plasminogen activator (uPA). RNA interference was used to down-regulate the expression of Notch1 and Notch3. Cell viability was assessed using MTT.

Results: Based on immunohistochemistry, high Notch1 expression was correlated with tumor size, tumor grade, metastasis, venous invasion and AJCC TNM stage. High Notch3 expression was only strongly correlated with metastasis, venous invasion and satellite lesions. Kaplan-Meier curves demonstrated that patients with high Notch1 or Notch3 expression were at a significantly increased risk for shortened survival time. In vitro, the down-regulation of Notch1 decreased the migration and invasion capacities of HCC cells by regulating CD44v6, E-cadherin, MMP-2, MMP-9, and uPA via the COX-2 and ERK1/2 pathways. Down-regulation of Notch3 only decreased the invasion capacity of HCC cells by regulating MMP-2 and MMP-9 via the ERK1/2 pathway.

Conclusions: Based on the migration and invasion of HCC, we hypothesize that targeting Notch1 may be more useful than Notch3 for designing novel preventive and therapeutic strategies for HCC in the near future.

Show MeSH
Related in: MedlinePlus