Related Experiment Video
Updated: May 24, 2026
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
Published on: July 3, 2013
Antitumor potential of SLPI promoter controlled recombinant caspase-3 expression in laryngeal carcinoma
1Department of Otorhinolaryngology, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
Abstract:
The purpose of this study is to develop a specific and efficient targeted gene therapy candidate approach for laryngeal carcinomas. Several promoters of human squamous cell carcinoma antigen 2(SCCA2), secretory leukocyte protease inhibitor (SLPI) and Survivin genes were cloned from human genomic DNA and evaluated for tumor-specific transcription potential in human laryngeal carcinoma Hep-2 cells by dual luciferase assays. One SLPI promoter fragment (677 bp) showed the highest efficiency and specificity, and was used to control the expression of a recombinant active caspases-3 (revCasp3), which could trigger apoptosis without activation of its upstream cascade elements once expressed in a cell, in an adenoviral vector (Ad-SLPI-revCasp3), and its antitumor efficacy was assessed. In vitro infection with Ad-SLPI-revCasp3 showed revCasp3 could be specifically expressed in Hep-2 cells, resulting in efficient activation of endogenous Caspase-3 and subsequent apoptosis of Hep-2 cells. In Hep-2 nude mice xenograft model, intratumoral administration of Ad-SLPI-revCasp3 significantly inhibited tumor growth without obvious loss of body weight and obvious hepatic toxicity. In summary, our study showed the specific and efficient apoptosis-inducing potential of Ad-SLPI-revCasp3, and this makes it a new candidate approach of targeted gene therapy for laryngeal squamous cell carcinoma, which needs further systematic investigation.
Insights
This study developed a targeted gene therapy using an adenoviral vector (Ad-SLPI-revCasp3) to specifically induce apoptosis in laryngeal carcinoma cells. The therapy showed significant tumor inhibition in mice with minimal toxicity, offering a promising new treatment candidate.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Laryngeal carcinomas present a significant challenge in cancer treatment.
- Targeted gene therapy offers a promising strategy for specific cancer cell destruction.
- Developing efficient and tumor-specific promoters is crucial for effective gene therapy vectors.
Purpose of the Study:
- To develop a specific and efficient targeted gene therapy candidate for laryngeal carcinomas.
- To identify and characterize tumor-specific promoters for controlling gene expression.
- To evaluate the antitumor efficacy of a novel adenoviral vector carrying a pro-apoptotic gene.
Main Methods:
- Cloning and evaluation of human squamous cell carcinoma antigen 2 (SCCA2), secretory leukocyte protease inhibitor (SLPI), and Survivin gene promoters.
- Dual luciferase assays to assess tumor-specific transcription potential in Hep-2 cells.
- Construction of an adenoviral vector (Ad-SLPI-revCasp3) expressing recombinant active caspases-3 (revCasp3) under SLPI promoter control.
- In vitro and in vivo assessment of Ad-SLPI-revCasp3 efficacy in laryngeal carcinoma models.
Main Results:
- A 677 bp SLPI promoter fragment demonstrated high efficiency and specificity for tumor transcription.
- Ad-SLPI-revCasp3 specifically expressed revCasp3 in Hep-2 cells, activating Caspase-3 and inducing apoptosis.
- Intratumoral administration of Ad-SLPI-revCasp3 significantly inhibited tumor growth in a Hep-2 nude mice xenograft model.
- The treatment showed no significant body weight loss or obvious hepatic toxicity.
Conclusions:
- Ad-SLPI-revCasp3 exhibits specific and efficient apoptosis-inducing potential against laryngeal carcinoma cells.
- This adenoviral vector represents a novel candidate for targeted gene therapy in laryngeal squamous cell carcinoma.
- Further systematic investigation is warranted to advance this therapeutic approach.