Lack of impact of the loss of constitutive folate receptor α expression, achieved by RNA interference, on the activity of the new generation antifolate pemetrexed in HeLa cells

Shrikanta Chattopadhyay, Yanhua Wang, Rongbao Zhao, I. David Goldman

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Pemetrexed [PMX (Alimta)] is a new generation antifolate with activity in a variety of solid tumors. It is an excellent substrate for most folate transporters, notably the reduced folate carrier (RFC) and folate receptor (FR)-α. The role of FR-α in PMX pharmacological activity is uncertain. Whereas high-level expression may enhance the activity of this agent, it is not clear what role constitutive levels of this transporter contribute to PMX activity. In this study, constitutive levels of FR-α expression were abolished by small interfering RNA-induced silencing in HeLa cells and RFC-null HeLa R5 cells as confirmed by Northern blotting, immunohistochemistry, and cell surface binding. PMX growth inhibition was unchanged in HeLa and R5 cells in the absence of FR-α expression. Loss of FR-α expression did not decrease net accumulation of PMX in either wild-type or RFC-null HeLa cells. Likewise, folate pools in wild-type HeLa cells were not decreased by FR-α gene silencing and were negligibly affected in the RFC-null R5 subline grown with 5-formyltetrahydrofolate. FR-α surface binding in HeLa cells was shown to be greater than that in a variety of other human solid tumor cell lines. Hence, constitutively expressed FR-α in HeLa cells does not contribute to PMX activity in the presence or absence of RFC function. This is likely the case in many human solid tumor cell lines.

Original languageEnglish (US)
Pages (from-to)7986-7993
Number of pages8
JournalClinical Cancer Research
Volume10
Issue number23
DOIs
StatePublished - Dec 1 2004

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Pemetrexed
Folic Acid Antagonists
RNA Interference
HeLa Cells
Folic Acid
Reduced Folate Carrier Protein
Null Lymphocytes
Tumor Cell Line
Folic Acid Transporters
Leucovorin
Gene Silencing
Northern Blotting
Small Interfering RNA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

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title = "Lack of impact of the loss of constitutive folate receptor α expression, achieved by RNA interference, on the activity of the new generation antifolate pemetrexed in HeLa cells",
abstract = "Pemetrexed [PMX (Alimta)] is a new generation antifolate with activity in a variety of solid tumors. It is an excellent substrate for most folate transporters, notably the reduced folate carrier (RFC) and folate receptor (FR)-α. The role of FR-α in PMX pharmacological activity is uncertain. Whereas high-level expression may enhance the activity of this agent, it is not clear what role constitutive levels of this transporter contribute to PMX activity. In this study, constitutive levels of FR-α expression were abolished by small interfering RNA-induced silencing in HeLa cells and RFC-null HeLa R5 cells as confirmed by Northern blotting, immunohistochemistry, and cell surface binding. PMX growth inhibition was unchanged in HeLa and R5 cells in the absence of FR-α expression. Loss of FR-α expression did not decrease net accumulation of PMX in either wild-type or RFC-null HeLa cells. Likewise, folate pools in wild-type HeLa cells were not decreased by FR-α gene silencing and were negligibly affected in the RFC-null R5 subline grown with 5-formyltetrahydrofolate. FR-α surface binding in HeLa cells was shown to be greater than that in a variety of other human solid tumor cell lines. Hence, constitutively expressed FR-α in HeLa cells does not contribute to PMX activity in the presence or absence of RFC function. This is likely the case in many human solid tumor cell lines.",
author = "Shrikanta Chattopadhyay and Yanhua Wang and Rongbao Zhao and Goldman, {I. David}",
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T1 - Lack of impact of the loss of constitutive folate receptor α expression, achieved by RNA interference, on the activity of the new generation antifolate pemetrexed in HeLa cells

AU - Chattopadhyay, Shrikanta

AU - Wang, Yanhua

AU - Zhao, Rongbao

AU - Goldman, I. David

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N2 - Pemetrexed [PMX (Alimta)] is a new generation antifolate with activity in a variety of solid tumors. It is an excellent substrate for most folate transporters, notably the reduced folate carrier (RFC) and folate receptor (FR)-α. The role of FR-α in PMX pharmacological activity is uncertain. Whereas high-level expression may enhance the activity of this agent, it is not clear what role constitutive levels of this transporter contribute to PMX activity. In this study, constitutive levels of FR-α expression were abolished by small interfering RNA-induced silencing in HeLa cells and RFC-null HeLa R5 cells as confirmed by Northern blotting, immunohistochemistry, and cell surface binding. PMX growth inhibition was unchanged in HeLa and R5 cells in the absence of FR-α expression. Loss of FR-α expression did not decrease net accumulation of PMX in either wild-type or RFC-null HeLa cells. Likewise, folate pools in wild-type HeLa cells were not decreased by FR-α gene silencing and were negligibly affected in the RFC-null R5 subline grown with 5-formyltetrahydrofolate. FR-α surface binding in HeLa cells was shown to be greater than that in a variety of other human solid tumor cell lines. Hence, constitutively expressed FR-α in HeLa cells does not contribute to PMX activity in the presence or absence of RFC function. This is likely the case in many human solid tumor cell lines.

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