腺病毒介导的FasL基因导入人胃癌细胞

Adenovirus-mediated FasL gene transfer into human gastric carcinoma.

作者信息

Zheng Shi-Ying, Li De-Chun, Zhang Zhi-De, Zhao Jun, Ge Jin-Feng

机构信息

Department of Cardiothoracic Surgery, The First Affiliated Hospital of Suzhou University, Suzhou 215006, Jiangsu Province, China.

出版信息

World J Gastroenterol. 2005 Jun 14;11(22):3446-50. doi: 10.3748/wjg.v11.i22.3446.

DOI:10.3748/wjg.v11.i22.3446

PMID:15948252

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4316001/

Abstract

AIM

To evaluate the possible value of FasL in gastric cancer gene therapy by investigating the effects of FasL expression on human gastric cancer cell line.

METHODS

An adenoviral vector encoding the full-length human FasL cDNA was constructed and used to infect a human gastric cancer (SGC-7901) cell line. FasL expression was confirmed by X-gal staining, flow cytometric analysis and RT-PCR. The effect of FasL on cell proliferation was determined by clonogenic assay, cytotoxicity was detected by MTT assay, and cell viability was measured by trypan blue exclusion. The therapeutic efficiency of Ad-FasL in vivo was investigated with a xenograft tumor model in nude mice.

RESULTS

SGC-7901 cells infected with Ad-FasL showed increased expression of FasL, resulting in significantly decreased cell growth and colony-forming activity when compared with control adenovirus-infected cells. The cytotoxicity of anti-Fas antibody (CH-11) in gastric cancer cells was stronger than that of ActD (91+/-8 vs 60+/-5, P<0.01), and the cytotoxicity of Ad-FasL was stronger than that of CH-11 (60+/-5 vs 50+/-2, P<0.05). In addition, G1-phase arrest (67.75+/-0.39 vs 58.03+/-2.16, P<0.05) and apoptosis were observed in Ad-FasL-infected SGC-7901 cells, and the growth of SGC-7901 xenografts in nude mice was retarded after intra-tumoral injection with Ad-FasL (54% vs 0%, P<0.0001).

CONCLUSION

Infection of human gastric carcinoma cells with Ad-FasL induces apoptosis, indicating that this target gene might be of potential value in gene therapy for gastric cancer.

摘要

目的

通过研究FasL表达对人胃癌细胞系的影响，评估FasL在胃癌基因治疗中的潜在价值。

方法

构建编码全长人FasL cDNA的腺病毒载体，用于感染人胃癌（SGC-7901）细胞系。通过X-gal染色、流式细胞术分析和RT-PCR确认FasL表达。通过克隆形成试验确定FasL对细胞增殖的影响，通过MTT试验检测细胞毒性，通过台盼蓝排斥法测量细胞活力。用裸鼠异种移植瘤模型研究Ad-FasL在体内的治疗效果。

结果

感染Ad-FasL的SGC-7901细胞显示FasL表达增加，与对照腺病毒感染的细胞相比，细胞生长和集落形成活性显著降低。抗Fas抗体（CH-11）对胃癌细胞的细胞毒性强于放线菌素D（91±8对60±5，P<0.01），Ad-FasL的细胞毒性强于CH-11（60±5对50±2，P<0.05）。此外，在感染Ad-FasL的SGC-7901细胞中观察到G1期阻滞（67.75±0.39对58.03±2.16，P<0.05）和凋亡，在裸鼠体内瘤内注射Ad-FasL后，SGC-7901异种移植瘤的生长受到抑制（54%对0%，P<0.0001）。