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目的探讨三氧化二砷(As2O3)和顺铂(DDP)对人大肠癌细胞株CCL-187端粒酶活性及细胞凋亡的影响。方法大肠癌细胞株CCL-187与空白培养液,含1.0μmol/LAs2O3、0.2μg/mLDDP、1.0μmol/LAs2O3合用0.2μg/mlDDP的培养液共同孵育1～6d。MTT法计算细胞生长抑制率,电镜下观察细胞形态学变化,RT-PCR法检测细胞端粒酶hTERT-mRNA的表达,TRAP-PCR-ELISA法检测细胞端粒酶活性。结果1.0μmol/LAs2O3能促进细胞分化,0.2μg/mLDDP和合用组均能诱导细胞凋亡,同时都下调端粒酶hTERT-mRNA的表达,降低端粒酶的活性,且具有时间依赖性。用药组与对照组相比差异有极显著性(P<0.01);合用组与单药组相比差异有极显著性(P<0.01)。细胞生长抑制率与细胞端粒酶hTERT-mRNA的表达、端粒酶活性的下调呈负相关。细胞端粒酶hTERT-mRNA的表达与细胞端粒酶活性呈正相关。结论1.0μmol/LAs2O3对人大肠癌CCL-187细胞有促进分化的作用,0.2μg/mLDDP和合用组有诱导凋亡的作用,其机制可能是下调细胞端粒酶hTERT-mRNA的基因表达,从而抑制端粒酶的活性。
Objective To investigate the effects of arsenic trioxide (As2O3) and cisplatin (DDP) on telomerase activity and apoptosis of human colorectal cancer cell line CCL-187. Methods Colorectal cancer cell line CCL-187 was incubated with blank medium, 1.0μmol / L As2O3, 0.2μg / mL DDP, 1.0μmol / L As2O3 and 0.2μg / ml DDP for 1 ~ 6 days. Cell growth inhibition rate was calculated by MTT method. Morphological changes were observed under electron microscope. Telomerase hTERT-mRNA expression was detected by RT-PCR and telomerase activity by TRAP-PCR-ELISA. Results 1.0μmol / L As2O3 could promote cell differentiation. Both 0.2μg / mL DDP and combination group could induce cell apoptosis and down-regulate the expression of telomerase hTERT-mRNA and decrease the telomerase activity in a time-dependent manner. There was a significant difference between the treatment group and the control group (P <0.01). The difference between the combination group and the single drug group was significant (P <0.01). Cell growth inhibition rate and telomerase hTERT-mRNA expression, telomerase activity was negatively correlated. The telomerase hTERT-mRNA expression was positively correlated with telomerase activity. Conclusions 1.0μmol / L As2O3 can promote the differentiation of human colorectal cancer cell line CCL-187, and 0.2μg / mL DDP combined with it can induce apoptosis. The possible mechanism is that the down-regulation of telomerase hTERT-mRNA gene expression Inhibit telomerase activity.