hTERT反义寡核苷酸对肝癌SMMC-7721细胞株细胞增殖和端粒酶活性的影响

目的研究hTERT基因反义寡核苷酸（ASPSODN）对SMMC-7721细胞端粒酶活性及细胞增殖的影响。方法不同浓度的ASPSODN转染到SMMC-7721细胞株24h和48h后分别采用MTT法、TRAP-ELISA法检测SMMC-7721细胞的增殖活性及端粒酶活性。结果各浓度组的ASPSODN均能降低端粒酶活性,作用24h时,不同浓度均明显受抑,其中0.8μmol/L ASPSODN较0.2μmol/L和0.4μmol/L组抑制作用更明显（P〈0.05）,但再增大浓度抑制作用并不随着增加。作用48h后,抑制作用减弱,低浓度组（0.2、0.4μmol/L）端粒酶活性恢复到正常水平,而高浓度组（0.8、1.0、1.2μmol/L）抑制作用依然明显,端粒酶活性低于60%。MTT法检测显示,不同浓度ASPSODN组对SMMC-7721细胞的生长均有抑制作用,随着浓度的增加,抑制作用明显,但随着时间的延长,抑制作用增加不明显。结论 ASPSODN靶向hTERT能特异性抑制SMMC-7721细胞增殖,明显下调端粒酶活性,hTERT可能成为肝癌治疗的一个靶点。

Effect of antisence oligodeoxynucleotide on cell proliferation and telomerase activity in SMMC-7721 cell line

Objective To observe the effect of hTERT anti-sense oligodenucletide（ASPSODN） on the telomerage activity and proliferation to human hepatic carcinoma SMMC-7721 cell line in vitro.Methods Human hepatic carcinoma SMMC-7721 cell was transfected with ASPSODN by liposome.The proliferation activity of SMMC-7721 cell line was determined by using methyl thiazolyl tetrazolium assay,and telomerase activity was detected by TRAP-ELISA 24h and 48h after treatment.Results The result shows telomerase activity was inhibited at different ASPSODN concentration,and 0.8 μmol/L ASPSODN is obvious Compared to 0.2μmol/L or 0.4μmol/L.But inhibiting effect did not increase when ASPSODN at 1.0 μmol/L or 1.2μmol/L compared with 0. 8μmol/L.However,inhibiting effect reduced after treatment 48 hours,and telomerase activity returned to normal level in 0.2μmol/ L and 0.4μmol/L group.But in the 0.8μmol/L、1.0μmol/L、1.2μmol/L groups,telomerase activity remain low.MTT showed proliferation of SMMC-7721 cell was suppressed by different ASPSODN concentration and The effects of suppression on cells rised with ASPSODN concentration increasing,but had not related with time increasing.Conclusion ASPSODN targeted hTERT can inhibit the proliferation of SMMC-7721 cell and decrease the telomerase activity.HTERT ati-sense may be a potential target of gene therapy for human liver cancer.