CAJ | 학술논문

目的研究COX-2反义RNA对肝癌细胞增殖抑制的作用及其机制,探讨肝癌治疗的新途径.方法人工合成的COX-2反义RNA片段及无关对照空质粒经脂质体包裹后作用CBRH7919细胞.通过MTT法、细胞周期分析、RT-PCR及裸鼠体内接种等方法测定细胞体内外增殖的变化.结果经COX-2反义RNA处理的CBRH7919细胞与对照组CBRH7919细胞相比,体外增殖速度减慢(细胞增殖抑制率达78%)、DNA合成受抑(S期细胞数为34.8%vs59.9%),细胞周期G_0/G.比例明显提高,裸鼠体内成瘤率下降(25%vs100%).而凋亡相关基因表达并无明显差异(P>0.05).结论 COX-2反义RNA可有效抑制肝癌细胞的体内外生长增殖,可用于实验性肿瘤基因治疗研究.
목적 연구COX-2반의RNA대간암세포증식억제적작용급기궤제,탐토간암치료적신도경.방법 인공합성적COX-2반의RNA편단급무관대조공질립경지질체포과후작용CBRH7919세포.통과MTT법、세포주기분석、RT-PCR급라서체내접충등방법측정세포체내외증식적변화.결과 경COX-2반의RNA처리적CBRH7919세포여대조조CBRH7919세포상비,체외증식속도감만(세포증식억제솔체78%)、DNA합성수억(S기세포수위34.8%vs59.9%),세포주기G_0/G.비례명현제고,라서체내성류솔하강(25%vs100%).이조망상관기인표체병무명현차이(P>0.05).결론 COX-2반의RNA가유효억제간암세포적체내외생장증식,가용우실험성종류기인치료연구.
Objective To construct a recombinant eukaryotic expression vector encoding rat COX-2 antisense RNA and investigate its effects on rat liver cancer cell proliferatiion. Methods The plasmid encoding anti-sense COX-2 was constructed by using cloning COX-2 cDNA fragment in the reverse direction into the pcDNA3. 1. Then the plasmid pcDNA3. l/COX-2as was transfered into rat hepatocacinoma cell line CBRH7919 with liposome and homologous recombination cell was named as CBRH7919-A. The cell proliferation, cell cycle and apoptosis were analyzed by MTT, flow cytometry and RT-PCR. CBRH7919 cells with or without pretreatment were inoculated into nude mice and the cell proliferation was observed in vivo. Results CBRH7919 treated with antisense COX-2 greatly inhibited the proliferation with a rate of 78% and DNA synthesis (PI of COX-2 group vs control group, 0. 361 vs 0. 784) of CBRH7919 cell line, decreased coloning formation in anchorage-independent assay. In vivo tumorigenic rate was greatly reduced in athymic nude mice as compared with untreated cell group (25% vs 100%). Apoptosis-related gene expression was not different as compared with untreated cell group (P>0. 05). Conclusion Antisense COX-2 RNA can inhibit the proliferation of rat liver cancer cell line in vitro as well as in vivo, suggesting that it has potential value in hepatocellular cancer gene therapy.