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Specific inhibition of tumor cells by oncogenic EGFR specific silencing by RNA interference.

ABSTRACT: Anticancer agents that have minimal effects on normal cells and tissues are ideal cancer drugs. Here, we show specific inhibition of human cancer cells carrying oncogenic mutations in the epidermal growth factor receptor (EGFR) gene by means of oncogenic allele-specific RNA interference (RNAi), both in vivo and in vitro. The allele-specific RNAi (ASP-RNAi) treatment did not affect normal cells or tissues that had no target oncogenic allele, whereas the suppression of a normal EGFR allele by a conventional in vivo RNAi caused adverse effects, i.e., normal EGFR is vital. Taken together, our current findings suggest that specific inhibition of oncogenic EGFR alleles without affecting the normal EGFR allele may provide a safe treatment approach for cancer patients and that ASP-RNAi treatment may be capable of becoming a safe and effective, anticancer treatment method.

SUBMITTER: Takahashi M

PROVIDER: S-EPMC3738543 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Specific inhibition of tumor cells by oncogenic EGFR specific silencing by RNA interference.

Takahashi Masaki M Chiyo Tomoko T Okada Takashi T Hohjoh Hirohiko H

PloS one 20130808 8

Anticancer agents that have minimal effects on normal cells and tissues are ideal cancer drugs. Here, we show specific inhibition of human cancer cells carrying oncogenic mutations in the epidermal growth factor receptor (EGFR) gene by means of oncogenic allele-specific RNA interference (RNAi), both in vivo and in vitro. The allele-specific RNAi (ASP-RNAi) treatment did not affect normal cells or tissues that had no target oncogenic allele, whereas the suppression of a normal EGFR allele by a co ...[more]

PMID: 23951344

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Project description:The objective of the present study was to investigate the expression levels of toll-like receptor 4 (TLR4) in glioma cells and the mechanisms underlying its regulatory effects on proliferation, migration and apoptosis of glioma cells. A total of three TLR4 silencing short hairpin (sh)RNA plasmids were established, and Lipofectamine® was used to the transfect the human glioma cell line U-87MG. Transfection efficiency was measured via flow cytometry. The interference plasmid exhibiting the largest silencing effect on TLR4 was screened for subsequent experiments using puromycin. Reverse transcription-quantitative PCR and western blot analysis were used to detect the TLR4 gene and protein expression levels, respectively, in stably transfected cells. Flow cytometry measured cell cycle and apoptosis and a wound healing assay was employed to assess the migration ability of transfected cells. The proliferation of transfected cells was detected using Cell Counting Kit-8 assay. TLR4-sh2 exhibited the highest transfection efficiency. Following transfection of U-87MG cells with TLR4-sh2 and negative control (NC) plasmids for 48 h and screening by puromycin, stable transfected cells were named U-87MG-Sh and U-87MG-NC cells respectively. The TLR4 gene and protein expression levels in the U-87MG-Sh cells were significantly lower than in U-87MG and U-87MG-NC cells. The apoptosis rate and the percentage of G0/1 cells were significantly higher, whereas the cell proliferation rate was notably lower, in U-87MG-Sh cells than in the U-87MG-NC and U-87MG cells. The proliferation rate and the cell migration ability of U-87MG-Sh cells were significantly lower than those of U-87MG-NC and U-87MG cells. TLR4 is associated with the proliferation of glioma cells. Inhibition of TLR4 expression levels significantly inhibited proliferation of glioma cells and induced apoptosis. The present study provided insights into the mechanisms associated with the development, progression and invasion ability of glioma cells.

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Specific inhibition of tumor cells by oncogenic EGFR specific silencing by RNA interference.

Publications

Specific inhibition of tumor cells by oncogenic EGFR specific silencing by RNA interference.

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