摘要
目的 探讨反义hTR基因转染对胃癌细胞系SGC 790 1恶性表型的逆转作用及其在肿瘤基因治疗中的价值。方法 构建包含端粒重复序列模板区的反义hTR基因真核表达载体 ,用脂质体法将其转染至胃癌细胞系SGC 790 1,比较观察反义hTR基因转染后 790 1细胞的hTRRNA表达、端粒酶活性、体外生长增殖特性和裸鼠体内致瘤能力的变化。结果 反义hTR基因转染的 790 1细胞hTRRNA表达和端粒酶活性明显降低、体外生长增殖能力降低、接触抑制恢复、细胞凋亡率增加、软琼脂集落形成能力和裸鼠体内致瘤能力完全消失。结论 反义hTR基因转染能使 790 1细胞的端粒酶活性明显下调并逆转其恶性表型 ,提示端粒酶是肿瘤治疗的较理想靶点 ,端粒酶抑制剂具有良好的临床应用前景。
Objective To study the effects of the transfection of antisense human telomerase RNA (hTR) on the malignant phenotype of gastric carcinoma cell line SGC 7901 and its potential role in the gene therapy for cancers. Methods An antisense hTR gene eukaryotic expression vector pCL ahTR including the template region of telomere repeats sequence was constructed with recombinant DNA technique and transfected into the cells of gastric carcinoma cell line SGC 7901 with liposome DOTAP. Then, the expression of hTR and antisense hTR was observed with RT PCR, telomerase activity with PCR ELISA, the cell morphology and cellular proliferation capacity with MTT assay and the efficiency of clone formation in soft agar, the cell cycle distribution and the apoptotic state with flow cytometry. The characteristics of the proliferation of SGC 7901 cells in vitro and the tumorigenecity in nude mice were also observed. Results The expression of hTR RNA, the activity of telomerase and the proliferative capacity of antisense hTR gene transfected SGC 7901 cells in vitro were obviously decreased and the apoptotic rate increased. The capability to form clones in soft agar and to develop carcinoma in nude mice totally vanished. Conclusion Our study demonstrated that after the transfection of antisense hTR gene, the telo merase activity of SGC 7901 cells was obviously down regulated and their malignant phenotype was reversed, which implies that telomerase is an ideal target in cancer therapy and telomerase inhibitors are promising agents in this aspect.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2000年第7期611-614,共4页
Journal of Third Military Medical University
基金
国家自然科学基金!资助项目 (39770 30 2 )
关键词
真核表达载体
基因治疗
胃肿瘤
反义hTR基因
human telomerase RNA components
telomerase
eukaryotic expression vector
antisense gene
gene therapy
gastric carcinoma
cell line
