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siRNA-mediated silencing of the 37/67-kDa high affinity laminin receptor in Hep3B cells induces apoptosis

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Abstract

The laminin-binding protein, variously called the 37/67-kDa high affinity laminin receptor or p40, mediates the attachment of normal cells to the laminin network, and also has a role as a ribosomal protein. Over-expression of this protein has been strongly correlated with the metastatic phenotype. However, few studies have investigated the cellular consequence of the ablation of this gene’s expression. To address this issue, the expression of the 37/67-kDa high affinity laminin receptor was knocked out with several siRNA constructs via RNA interference in transformed liver (Hep3B) cells. In each case where the message was specifically ablated, apoptosis was induced, as determined by annexin V/propidium iodide staining, and by double staining with annexin V and an antibody directed against the 37/67-kDa high affinity laminin receptor. These results suggest that this protein plays a critical role in maintaining cell viability.

Abbreviations

GAPDH:

glyceraldehyde 3-phosphate dehydrogenase

LBP-p40:

laminin-binding protein precursor p40

LRP:

laminin receptor precursor

PrPc :

cellular prion protein

siRNA:

small interfering RNAs

37LBP:

37-kDa laminin binding protein

67LR:

67-kDa laminin receptor

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Correspondence to Duncan R. Smith.

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Key words

  • siRNA
  • RNA interference
  • Laminin receptor
  • p40
  • Ribosomal
  • Liver
  • Silencing
  • LAMR1