Improved therapeutic effectiveness by combining recombinant p14ARF with antisense complementary DNA of EGFR in laryngeal squamous cell carcinoma

Abstract

PurposeThe tumor suppressor p14ARF and proto-oncogene epidermal growth factor receptor (EGFR) play important roles in the development of laryngeal squamous cell carcinoma (LSCC). This study was aimed to determine whether combining recombinant p14ARF with antisense complementary DNA of EGFR could improve the therapeutic effectiveness in LSCC. Materials and methodsAfter human larynx cancer cells (Hep-2) were infected with recombinant adenoviruses (Ad-p14ARF and Ad-antisense EGFR) together or alone in vitro, the proliferation and cell cycle distribution of Hep-2 cells were detected by MTT assay and flow cytometer analysis, respectively. Furthermore, the antitumor effects of recombinant adenoviruses together or alone on Hep-2 xenografts were examined in vivo. The levels of p14ARF and EGFR expressed in Hep-2 cells and xenografts were determined by western blot assay. ResultsAd-p14ARF combining with Ad-antisense EGFR markedly inhibited the Hep-2 proliferation compared with alone (P=0.001, P=0.002 respectively). Combination of Ad-p14ARF and Ad-antisense EGFR led to the proportion of Hep-2 cells in G0/G1 phases increased by up to 86.9%. The down-expression of EGFR protein and overexpression of p14ARF protein were observed in vitro and in vivo, and this effect was preserved when Ad-p14ARF was combined with Ad-antisense EGFR. Besides, Ad-p14ARF plus Ad-antisense EGFR significantly (P<0.05) increased the antitumor activity against Hep-2 tumor xenografts comparing with Ad-p14ARF or Ad-antisense EGFR alone. ConclusionCombination Ad-p14ARF with Ad-antisense EGFR significantly increased the antitumor responses in LSCC. An effectively potential gene therapy to prevent proliferation of LSCC was provided.