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Experimental gene therapy using p21Waf1 gene for esophageal squamous cell carcinoma by gene gun technology
Yuichi Tanaka1, Teruhiko Fujii, Hideaki Yamana
1Department of Surgery, Kurume University School of Medicine, 67 Asahi-machi, Kurume, Fukuoka 830-0011, Japan.
Abstract:
In our previous study, the proliferation rate of esophageal squamous cell carcinoma cell lines, which poorly expressed p21Waf1, was found to be regulated by p21Waf1 gene transfection using adenovirus vector. In the present study, in order to examine the effect of p21Waf1 gene therapy in esophageal cancer, we used gene gun technology, which proved to be a powerful method to introduce the p21Waf1 gene into esophageal cancer cells. p21Waf1 transfection to KE3 and YES2 cells (weakly expressed p21Waf1 protein cells) showed a high expression of p21Waf1 protein after applying this gene gun technique. In KE3 and YES2 cells, statistical significant growth inhibition was observed after p21Waf1 transfection compared with LacZ transfection (KE3, p=0.0009; YES2, p<0.0001). In in vivo transfection experiments, on day 14, the estimated volume of KE3 tumors subjected to p21Waf1 gene transfection was 95% in comparison with the pretreatment volume on day 0, while the volume of KE3 tumors subjected to LacZ gene therapy increased to 268%. On day 14, the estimated volume of YES2 tumors subjected to either p21Waf1 or LacZ gene therapy increased to 474 and 686%, respectively. In KE3 and YES2 cells, significant growth inhibition was observed after combination therapy using p21Waf1 transfection and anticancer drug 5-fluorouracil (5Fu) compared with 5Fu alone (KE3, p<0.0001; YES2, p<0.0001). In conclusion, p21Waf1 gene therapy using the gene gun technique significantly inhibited the low basal p21Waf1 expressed esophageal cancer cell growth in vitro and in vivo. Furthermore, p21Waf1 transfection strongly enhanced the effect of 5Fu suggesting that p21Waf1 may prove beneficial in chemotherapy combined with gene therapy using gene gun technology in patients with esophageal cancer who have a low level of p21Waf1 expressed tumor.
Insights
Gene gun delivery of the p21Waf1 gene effectively inhibits esophageal cancer cell growth in vitro and in vivo. This p21Waf1 gene therapy also enhances chemotherapy, offering a promising treatment for esophageal cancer.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- Esophageal squamous cell carcinoma (ESCC) often exhibits low expression of the p21Waf1 protein, a key regulator of cell proliferation.
- Previous studies indicated that p21Waf1 gene transfection could regulate ESCC proliferation.
- Gene gun technology offers a novel method for efficient gene delivery into cancer cells.
Purpose of the Study:
- To investigate the efficacy of p21Waf1 gene therapy in esophageal cancer using gene gun technology.
- To evaluate the in vitro and in vivo effects of p21Waf1 gene transfection on ESCC cell growth.
- To assess the synergistic effect of p21Waf1 gene therapy combined with 5-fluorouracil (5Fu) chemotherapy.
Main Methods:
- p21Waf1 gene was introduced into esophageal cancer cell lines (KE3 and YES2) with low p21Waf1 expression using gene gun technology.
- In vitro and in vivo transfection experiments were conducted to assess tumor growth inhibition.
- Combination therapy involving p21Waf1 transfection and 5-fluorouracil (5Fu) was evaluated.
Main Results:
- Gene gun delivery resulted in high p21Waf1 protein expression in KE3 and YES2 cells.
- Significant growth inhibition of esophageal cancer cells was observed in vitro following p21Waf1 transfection.
- In vivo studies demonstrated substantial tumor volume reduction in p21Waf1-treated KE3 xenografts compared to controls.
- Combination therapy with p21Waf1 gene therapy and 5Fu significantly enhanced anti-cancer effects compared to 5Fu alone.
Conclusions:
- p21Waf1 gene therapy utilizing gene gun technology is effective in inhibiting the growth of esophageal cancer cells with low p21Waf1 expression, both in vitro and in vivo.
- p21Waf1 gene transfection potentiates the efficacy of 5-fluorouracil chemotherapy.
- This combined gene and chemotherapy approach holds promise for treating esophageal cancer patients with low p21Waf1 tumor expression.
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