Targeted knockdown of notchl inhibits invasion of human prostate cancer cells concomitant with inhibition of matrix metalloproteinase-9 and urokinase plasminogen activator

Bilal Hafeez, Vaqar Mustafa Adhami, Mohammad Asim, Imtiaz A. Siddiqui, Kumar M. Bhat, Weixiong Zhong, Mohammad Saleem, Maria Din, Vijayasaradhi Setaluri, Hasan Mukhtar

Research output: Contribution to journalArticle

110 Citations (Scopus)

Abstract

Purpose: Notch, a type 1 transmembrane protein, plays a key role in the development of many tissues and organ types. Aberrant Notch signaling, found in a wide variety of human cancers, contributes to tumor development. Because Notchl was found to be overexpressed in prostate cancer (PCa) cells and human PCa tissue, we therefore tested our hypothesis that overexpression of Notchl in PCa promotes tumor invasion. Experimental Design: Notchl expression was evaluated in human PCa cells and human PCa tissues. PCa cells were transiently transfected with Notchl-specific small interfering RNAs in concentrations ranging from 30 to 120 nmol/L and subsequently evaluated for effects on invasion and expression analysis for molecules involved in invasion. Results: Small interfering RNA-mediated knockdown of Notchl in PC3 and 22Rv1 PCa cells dramatically decreased their invasion. Focused cDNA array revealed that Notchl knockdown resulted in significant reduction in the expression of urokinase plasminogen activator (uPA) and matrix metalloproteinase-9 (MMP9) gene transcripts. These data were further verified by reverse transcription-PCR, real-time reverse transcription-PCR, and immunoblot analysis. Knockdown of Notchl was also observed to significantly reduce the mRNA expression and protein levels of uPA and its receptor uPAR. A significant reduction in MMP9 expression in Notchl knockdown cells suggested a role for Notchl in augmenting MMP9 transcription. Conclusions: Our data show the involvement of Notchl in human PCa invasion and that silencing of Notchl inhibits invasion of human PCa cells by inhibiting the expression of MMP9 and uPA. Thus, targeting of Notchl could be an effective therapeutic approach against PCa.

Original languageEnglish (US)
Pages (from-to)452-459
Number of pages8
JournalClinical Cancer Research
Volume15
Issue number2
DOIs
StatePublished - Jan 15 2009

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Plasminogen Activators
Matrix Metalloproteinase 9
Urokinase-Type Plasminogen Activator
Prostatic Neoplasms
Small Interfering RNA
Reverse Transcription
Urokinase Plasminogen Activator Receptors
Neoplasms
Oligonucleotide Array Sequence Analysis
Real-Time Polymerase Chain Reaction
Proteins
Research Design
Polymerase Chain Reaction
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

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Targeted knockdown of notchl inhibits invasion of human prostate cancer cells concomitant with inhibition of matrix metalloproteinase-9 and urokinase plasminogen activator. / Hafeez, Bilal; Adhami, Vaqar Mustafa; Asim, Mohammad; Siddiqui, Imtiaz A.; Bhat, Kumar M.; Zhong, Weixiong; Saleem, Mohammad; Din, Maria; Setaluri, Vijayasaradhi; Mukhtar, Hasan.

In: Clinical Cancer Research, Vol. 15, No. 2, 15.01.2009, p. 452-459.

Research output: Contribution to journalArticle

Hafeez, Bilal ; Adhami, Vaqar Mustafa ; Asim, Mohammad ; Siddiqui, Imtiaz A. ; Bhat, Kumar M. ; Zhong, Weixiong ; Saleem, Mohammad ; Din, Maria ; Setaluri, Vijayasaradhi ; Mukhtar, Hasan. / Targeted knockdown of notchl inhibits invasion of human prostate cancer cells concomitant with inhibition of matrix metalloproteinase-9 and urokinase plasminogen activator. In: Clinical Cancer Research. 2009 ; Vol. 15, No. 2. pp. 452-459.
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AU - Adhami, Vaqar Mustafa

AU - Asim, Mohammad

AU - Siddiqui, Imtiaz A.

AU - Bhat, Kumar M.

AU - Zhong, Weixiong

AU - Saleem, Mohammad

AU - Din, Maria

AU - Setaluri, Vijayasaradhi

AU - Mukhtar, Hasan

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N2 - Purpose: Notch, a type 1 transmembrane protein, plays a key role in the development of many tissues and organ types. Aberrant Notch signaling, found in a wide variety of human cancers, contributes to tumor development. Because Notchl was found to be overexpressed in prostate cancer (PCa) cells and human PCa tissue, we therefore tested our hypothesis that overexpression of Notchl in PCa promotes tumor invasion. Experimental Design: Notchl expression was evaluated in human PCa cells and human PCa tissues. PCa cells were transiently transfected with Notchl-specific small interfering RNAs in concentrations ranging from 30 to 120 nmol/L and subsequently evaluated for effects on invasion and expression analysis for molecules involved in invasion. Results: Small interfering RNA-mediated knockdown of Notchl in PC3 and 22Rv1 PCa cells dramatically decreased their invasion. Focused cDNA array revealed that Notchl knockdown resulted in significant reduction in the expression of urokinase plasminogen activator (uPA) and matrix metalloproteinase-9 (MMP9) gene transcripts. These data were further verified by reverse transcription-PCR, real-time reverse transcription-PCR, and immunoblot analysis. Knockdown of Notchl was also observed to significantly reduce the mRNA expression and protein levels of uPA and its receptor uPAR. A significant reduction in MMP9 expression in Notchl knockdown cells suggested a role for Notchl in augmenting MMP9 transcription. Conclusions: Our data show the involvement of Notchl in human PCa invasion and that silencing of Notchl inhibits invasion of human PCa cells by inhibiting the expression of MMP9 and uPA. Thus, targeting of Notchl could be an effective therapeutic approach against PCa.

AB - Purpose: Notch, a type 1 transmembrane protein, plays a key role in the development of many tissues and organ types. Aberrant Notch signaling, found in a wide variety of human cancers, contributes to tumor development. Because Notchl was found to be overexpressed in prostate cancer (PCa) cells and human PCa tissue, we therefore tested our hypothesis that overexpression of Notchl in PCa promotes tumor invasion. Experimental Design: Notchl expression was evaluated in human PCa cells and human PCa tissues. PCa cells were transiently transfected with Notchl-specific small interfering RNAs in concentrations ranging from 30 to 120 nmol/L and subsequently evaluated for effects on invasion and expression analysis for molecules involved in invasion. Results: Small interfering RNA-mediated knockdown of Notchl in PC3 and 22Rv1 PCa cells dramatically decreased their invasion. Focused cDNA array revealed that Notchl knockdown resulted in significant reduction in the expression of urokinase plasminogen activator (uPA) and matrix metalloproteinase-9 (MMP9) gene transcripts. These data were further verified by reverse transcription-PCR, real-time reverse transcription-PCR, and immunoblot analysis. Knockdown of Notchl was also observed to significantly reduce the mRNA expression and protein levels of uPA and its receptor uPAR. A significant reduction in MMP9 expression in Notchl knockdown cells suggested a role for Notchl in augmenting MMP9 transcription. Conclusions: Our data show the involvement of Notchl in human PCa invasion and that silencing of Notchl inhibits invasion of human PCa cells by inhibiting the expression of MMP9 and uPA. Thus, targeting of Notchl could be an effective therapeutic approach against PCa.

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