Suppression of Survivin Expression by Short Hairpin RNA Induces Apoptosis in Human Laryngeal Carcinoma Cells

Abstract

Purpose: The aim of this study was to evaluate the suppression effect of survivin shRNA on the expression of the survivin gene in the human laryngeal cancer cell line Hep-2. Procedures: 60 cases of laryngeal squamous-cell carcinoma (LSCC) and 10 cases of normal laryngeal mucosa were examined using immunohistochemistry to determine whether the expression of survivin correlated with tumorigenesis. Three plasmid vectors of short hairpin RNA (shRNA) specific for survivin were designed and generated. Western blot and real-time PCR analysis of survivin expression in Hep-2 cells was performed 48 h after transfection. The growth curve was used to determine the cell proliferation. Propidium iodide (PI) single staining was applied to detect the cell cycle. The apoptosis of the cells was analyzed by flow cytometry with the FITC-annexin-V/PI double staining and PI single staining. Results: 68.33% (41 out of 60) of tumors were positive for survivin expression and significantly associated with lymph node metastasis and advanced stage. In contrast, no expression of survivin in normal mucosa was detected. Transfection of Hep-2 cells with survivin shRNA significantly inhibited survivin expression at both the mRNA and the protein level in Hep-2 cells. Downregulation of survivin resulted in increasing the apoptosis index, but the results showed no obvious influence on cell cycle. Conclusions: This study demonstrates that survivin shRNA effectively inhibits survivin gene expression in Hep-2 cells leading to growth suppression and apoptotic induction in Hep-2 cells.

Survivin and laryngeal carcinoma prognosis: nuclear localization and expression of splice variants

“…Many preliminary lines of evidence indicate that survivin inhibition by gene therapy (antisense oligonucleotides) and by small molecule inhibitors in head and neck carcinoma cell lines increases the anti‐tumour activity of several cytotoxic and other targeted therapies significantly 5 . Only a very limited number of the reported experiences of survivin inhibition have focused on LSCC cell lines, however 32–34 …”

Effect of apoptotic and proliferative indices, P-glycoprotein and survivin expression on prognosis in laryngeal squamous cell carcinoma

“…Moreover, survivin overexpression is usually correlated with low apoptotic and high proliferative activities, which favors the survival of cancer cells [17]. Previously, several in vitro studies have demonstrated that inhibition of survivin gene expression could lead to apoptotic induction and growth suppression in human laryngeal carcinoma cells [29,30]. To further determine whether survivin contributes to the apoptotic inhibition and proliferative promotion in laryngeal cancer, we investigated the survivin expression in all LSCCs evaluated.…”

Survivin-Based Treatment Strategies for Squamous Cell Carcinoma

“…The same results were achieved by transfecting a plasmid-containing survivin shRNA into an OSCC tongue cell line: shRNA inhibited the expression of survivin at both the protein and mRNA level, inhibited cell proliferation via caspase-3 activity induction, and enhanced chemosensitivity to cisplatin [ 115 ]. Survivin shRNA-mediated knockdown suppressed tumor growth and induced apoptosis after transfection into human laryngeal carcinoma cell lines [ 116 ]. Ngan et al [ 117 ] evaluated the responses to oxaliplatin, a third-generation platinum-based chemotherapeutic agent, in two esophageal cancer cell lines, one derived from SCC and one from adenocarcinoma.…”