survivin及bcl-2基因表达在胃癌细胞株MGC803顺铂耐药中的作用

The Affection of survivin and bcl-2 mRNA Expression on Drug- resistance of Gastric Cancer Cell Line MGC803 to Cisplatin

目的探讨胃癌细胞株MGC803对顺铂(CDDP)耐药产生的有关机制。方法观察不同浓度CDDP(0.1、1.0、10.0mg/L)对MGC803细胞增殖、凋亡及抗凋亡基因survivin,bcl-2mRNA表达的影响,用MTT法检测细胞生长抑制率;荧光染色检测细胞凋亡率;流式细胞仪测定细胞周期的变化;RT-PCR检测survivin、bcl-2mRNA的表达。结果10mg/L的CDDP对MGC803细胞有较强的抑制增殖、促进凋亡作用,而0.1、1.0mg/L的CDDP干预24h后,其抑制细胞增殖,促进细胞凋亡的作用逐渐消失;CDDP作用于细胞48h后,细胞S期增加,G2/M期下降;MGC803细胞在1.0mg/L的CDDP作用下,survivinmRNA表达自24h后逐渐增高,bcl-2mRNA表达逐渐下降(P<0.05)。结论CDDP可干扰MGC803细胞周期,但该细胞对低浓度CDDP易于产生耐药,survivinmRNA表达增高可能是MGC803细胞对CDDP产生耐药原因之一。

Objective To investigate the drug-resistance mechanisms of gastric cancer cell to cisplatin （CDDP）. Methods Gastric cancer cell line MGC803 were treated by CDDP（0. 1, 1.0, 10.0mg/L） for 12, 24, 48 and 72hours. Proliferation was detected by MTT assay and apoptosis was measured by fluorescence staining. Cell cycle was detected by flow cytometry, survivin and bcl-2 mRNA expression were detected by RT-PCR Results High concentration of CDDP（10. 0mg/L） could inhibit proliferation and induce apoptosis, hut low concentration of CDDP（0. 1, 1.0mg/L） did not. the effects of inhibiting proliferation and inducing apoptosis did not increase significantly after 24 hours, The cell cycle was altered after the cells were treated by CDDP. S phase of cells were increased and G2/M phase of cells were decreased, survivin mRNA expression increased and bcl-2 mRNA expression decreased gradually after treated by CDDP（1.0mg/L） for 24h（P〈0. 05）. Conclusion CDDP could arrest the cell cycle on MGC803 cell. But the cells were easily developing drug-resistance to low concentration of CDDP. Over expression of survivin mRNA may be one of the mechanisms of drug resistance in MGC803 cells.