摘要
目的:探讨棘霉素诱导肝癌SMMC7721细胞的凋亡作用及对转录因子Twist、p53、Sur-vivin和hTERT(人端粒酶)基因的影响。方法:采用MTT法测定棘霉素对肝癌细胞的生长抑制率;半定量逆转录聚合酶链反应(RT-PCR)检测与肝癌的发生和转移密切相关的转录因子Twist,与细胞增殖和凋亡联系紧密的p53、Sur-vivin和hTERT的mRNA表达水平的改变;利用Western blot方法分析棘霉素治疗后上述基因的蛋白表达情况。结果:体外实验证实,棘霉素浓度>10.0ng/mL各组对细胞的生长抑制均显著增加,P<0.05,随棘霉素药物浓度增加,细胞抑制率明显增大。棘霉素抑制肝癌细胞株SMMC7721的作用呈剂量-时间依赖性。RT-PCR方法证实,随着棘霉素剂量的增加,肝癌SMMC7721细胞株中Twist、Survivin和hTERTmRNA表达水平逐渐下调,P<0.01,p53mRNA表达水平微弱上调,P<0.05。Western blot检测显示,Twist、Survivin和hTERT蛋白的表达量呈递减趋势,P<0.01,p53蛋白的表达微弱上调,P<0.01。结论:随着棘霉素剂量的增加,细胞凋亡率显著增加。棘霉素能够抑制与肝癌生长密切相关基因的Twist、Survivin和hTERTmRNA表达,促进p53mRNA表达上调,相应蛋白的表达呈下调和上调。棘霉素可能通过抑制肝癌细胞的生长、转移和促进肝癌细胞的凋亡起到抑制肿瘤生长的作用。
OBJECTIVE：To study the effect of apoptosis inducing human hepatocellular carcinoma （SMMC7721） cells by echinomycin and observe the expressions of Twist, p53, Survivin hTERT mRNA in SMMC7721 cells treated by echinomycin. METHODS： The effects of echinomycin and its concentration on the cell growth were examined by methyltetrazolium （MTT） assay; the levels of Twist, p53, hTERT and Survivin mRNA were measured by reverse transcriptionpolymerase chain reaction （RT-PCR）; the expressions of Twist, p53, Survivin and the hTERT protein were detected with Western blot analysis. RESULTS.. When echinomycin concentration was more than 10.0 ng/mL, the cell growth inhibition was significantly increased （P〈0.05）. Cell inhibitory rate was increased with augment of concentration of echinomycin in vitro. Echinomycin inhibited SMMC7721 cell at a time and dose dependent manner. Both the detecting of RT-PCR and Western blot showed that the expressions of Twist, Survivin and hTERT were down-regulated, respectively, P〈0.01 but p53 was weak up-regulated, P〈0.05. CONCLUSIONS： The cell inhibitory rate is evaluated with the increasing dose of echinomycin. The levels of Twist, Survivin, hTERT are down-regulated when the SMMC7721 cells are treated with ehinomycin, but contrary p53. Meanwhile, the corresponding protein expression is lowered and raised. Through inducing apoptosis and inhibiting proliferation and metastasis of cancer cells, echinomycin could be considered to inhibit the growth of tumor cells.
出处
《中华肿瘤防治杂志》
CAS
2008年第2期105-108,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
广西科技厅自然基金资助项目(桂科自0640121
桂科能0630006-5E3Z)
