NF-кB反义寡核苷酸对胃癌细胞裸鼠体内成瘤性及血管新生的影响

Effect of NF-кB antisense oligonucleotides on tumorigenesis and angiogenesis of gastric cancer cells in nude mice

目的:研究核转录因子(nuclear transcription factorκB,NF-κB)反义寡核苷酸(antisense oligodeoxynucleotide,ASODN)转染人胃癌细胞SGC-7901后,对该细胞裸鼠体内致瘤性和血管新生的影响。方法:设计合成NF-κB基因激活亚基P65启动子区ASODN,转染胃癌SGC-7901细胞株,利用荧光染色法观察转染效果,MTT法检测细胞的体外生长抑制率。建立裸鼠荷瘤模型,观察该细胞裸鼠皮下成瘤生长曲线变化,利用HE染色和TUNEL法检测肿瘤微血管密度(microvessel density,MVD)和凋亡指数(apoptotic index,AI)。结果:经P65ASODN转染的SGC-7901细胞体外增殖反应明显受到抑制(P<0.05)。体内成瘤实验结果显示,转染P65ASODN的SGC-7901细胞荷瘤裸鼠肿瘤体积明显小于其他各组(P<0.05),肿瘤组织的AI明显高于其他各组(P<0.01),肿瘤组织中的MVD明显少于其他各组(P<0.01)。结论:转染NF-κB/P65ASODN可明显抑制SGC-7901细胞的体内外增殖,该现象可能与P65ASODN诱导细胞凋亡,抑制肿瘤组织血管新生有关。

Objective：To investigate the effects of transfection of antisense oligodeoxynucleotide （ASODN） of nuclear transcription factor κB （NF-κB） on tumorigenesis and angiogenesis of human gastric cancer cell line SGC7901 in nude mice. Methods：ASODN targeted P65 promoter region of NF-κB was designed, synthesized, and transfected into gastric cancer cell line SGC-7901. Fluorescent staining was used to observe transfection efficacy. The growth-inhibiting rate of cells was measured by MTT assay in vitro. The subcutaneous xenograft model of human gastric cancer cells was established in nude mice and the tumor growth curve was observed. The microvessel density （MVD） was detected by HE staining, and the apoptotic index （AI） was quantitatively determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling （TUNEL）. Results：The cell proliferation was significantly inhibited in P65 ASODN-transfected group in vitro （ P 〈 0.05 ）. In vivo tumor formation test showed that the tumor volume in nude mice in ASODN group was obviously smaller than that of other groups （ P 〈 0.05 ） and the AI was significantly higher than that of other groups （P〈0.001）. The MVD in ASODN group was markedly lower than that of other groups （P 〈0.01 ）. Conclusion： Transfection of P65 ASODN significantly inhibited the proliferation of SGC-7901 cells in vitro. The mechanism may be related to induction of cell apoptosis and inhibition of angiogenesis causedby 1＇65 ASODN.