反义hTR对人胃癌细胞株SGC-7901恶性表型的逆转作用

目的 探讨反义ｈＴＲ基因转染对胃癌细胞系ＳＧＣ－７００１恶性有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率胃癌细胞系ＳＧＣ－７９０１,比较观察反义ｈＴＲ基因转染后７９０１细胞的ｈＴＲＲＮＡ表达、端粒酶活性、体外生长增殖降低,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

Antisense human telomerase RNA reverses malignant phenotype of gastric carcinoma cell line SGC-7901

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.