Small interfering RNA (siRNA) inhibits the expression of the Her2/neu gene, upregulates HLA class I and induces apoptosis of Her2/neu positive tumor cell lines

Abstract

Silencing of a specific mRNA using double stranded RNA oligonucleotides represents one of the newest technologies for suppressing a specific gene product. Small interfering RNA (siRNA) are 21 nucleotides long, double stranded RNA fragments that are identical in sequence to the target mRNA. We designed 3 such siRNA against the Her2/neu (

The oncogene HER2: its signaling and transforming functions and its role in human cancer pathogenesis

“…A number of studies using antisense, ribozyme or small interfering RNA (siRNA) methodologies to suppress HER2 expression in human cancer cell lines consistently show that HER2-overexpressing tumor cells are dependent on HER2 and undergo growth inhibition and apoptosis in cell culture, or tumor regression in vivo, in the absence of HER2 expression, while tumor types that do not overexpress HER2 are not sensitive to HER2 knockdown (Colomer et al, 1994;Juhl et al, 1997;Roh et al, 2000;Choudhury et al, 2004;Faltus et al, 2004). A kinasedead mutant of activated HER2 competes with and reverses the transformed phenotype induced by activated HER2 (Messerle et al, 1994).…”

HER-2/neu Mediates Oncogenic Transformation via Altered CREB Expression and Function

“…Then, cells were transfected with 1.5 mg/well ScaI-digested murine CREB1-specific (for NIH3T3 and HER-2/neu þ cells) or HER-2/neu-specific (MCF-7 cells) shRNA-encoding plasmid (SABioscience) using PolyFect (Qiagen) according to the manufacturer's instructions. A nonsense (NC) construct served as the control as recently described (22). Twenty-four hours after transfection, puromycin-resistant (pur R ) colonies were selected in medium supplemented with 3 mg puromycin/mL.…”

Targeting the function of the HER2 oncogene in human cancer therapeutics

“…Experimental models of HER2-overexpressing cancer cells using antisense, ribozyme or short interfering RNA (siRNA) methodologies consistently show that HER2 knockdown induces apoptosis in cell culture, or tumor regression in vivo, in the absence of HER2 expression, while tumor types that do not overexpress HER2 are not sensitive to HER2 knockdown (Colomer et al, 1994;Juhl et al, 1997;Roh et al, 2000;Choudhury et al, 2004;Faltus et al, 2004). Similar results are seen with kinase-dead HER2 and intracellular single-chain anti-HER2 antibodies (Beerli et al, 1994;Messerle et al, 1994;Deshane et al, 1996).…”