摘要
目的 研究特定序列人端粒酶反义寡核苷酸(AS -ODN)抑制三种分化程度不同的胃癌细胞(MKN -4 5、SGC -790 1、MKN -2 8)生长的可能性,并阐述其抑制胃癌细胞生长的作用与胃癌细胞分化程度的相关性。方法 在指定的作用时间和浓度等条件下,以AS -ODN作用于不同分化程度的三种胃癌细胞,用改良端粒酶活性定量检测法测定AS -ODN片段作用前后胃癌细胞的端粒酶活性;用锥虫蓝染色法观察细胞活力;用倒置显微镜、电镜、流式细胞仪和原位末端标记(TUNEL)法观察细胞凋亡情况。结果 以指定浓度的AS- ODN作用后,MKN- 4 5和SGC -790 1细胞出现明显的端粒酶活性和细胞生长抑制(P <0 .0 5 ) ,但在同样浓度条件下,MKN -2 8胃癌细胞只出现端粒酶活性抑制。错义序列对照组则无明显变化。以10 μmol/L的AS- ODN连续作用三种胃癌细胞96h后,光镜、电镜和TUNEL法检测均发现MKN- 4 5和SGC- 790 1细胞表现出特有的凋亡征象,流式细胞仪检测证实MKN -4 5和SGC- 790 1细胞的平均凋亡率在4 4 .75 %和33.5 6 % ,错义序列对照组则无明显变化(P <0 .0 5 )。结论 AS ODN能有效抑制胃癌细胞生长,其作用机制主要是抑制端粒酶活性和诱导细胞凋亡。AS -ODN对中分化胃癌细胞的生长抑制最显著,对低分化胃癌细胞的抑制作用略强于高分化胃癌细胞,?
Objective To investigate the effect of specialized human tel omerase antisense oligodeoxyribonucleotides (AS-ODN) on the growth inhibition o f well, moderate and poor differentiated gastric cancer cell lines, and to explore its inhibitory mechanism and the correlation between the inhibition ratio of gastric cancer cells and differentiation of the tumor cells. Methods Under the given circumstances, three distinct differentiated gastric cancer cell lines were treated with AS-ODN. The telomerase a ctivities were measured by modified telomeric repeat amplification protocal assa y. The cell viability was detected by Trypan blue test, and the cell apoptosis was determined by cell morphological observation under light and electromicroscope, flow cytometry an d TUNEL assay. Results The telomerase activity and cell growth were apparently suppressed in MK N45 and SGC7901 cells, under defined concentrations of AS-ODN. Whilst, in MKN28 cells, only telomerase activity was suppressed at same concentration . There were no obvious changes in non-antisense oligomers treated group. The apoptotic features of MKN45 and SGC7901 were noticed by microscopic observa tion, TUNEL assay, after three distinct gastric cancer cell lines being continuo usly exposed to 10 μmol/L AS-ODN for 96 h. Furthermore, the flow cytometric analy sis verified that the average apoptotic rate of MKN45 and SGC7901 was 44.75% and 33.56% respectively, but there were no obvious changes in non-antisense oli gomers treated group (P< 0.05). Conclusions AS-ODN inhibit the growth of gastric cancer cells und er the mechanism involved in the inhibition of telomerase activitity and induction of cell apopt osis.The inhibitory effects of AS-ODN were relatively strong in moderately differentiated, moderate in poor differe ntiated and weak in well differentiated ones. It indicates that the mechanism of AS-ODN is not only dependents on the sequence specialty of the antisense oligomers, but a lso depends on the biological characters of gastric cancer cell lines.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2004年第11期663-667,共5页
Chinese Journal of Digestion
