顺铂诱导人鼻咽癌CNE2细胞衰老及其相关基因表达谱的研究

目的 探讨顺铂体外诱导人鼻咽癌CNE2细胞衰老的效应及其分子机制。方法 采用 MTT 比色法观察顺铂对CNE2细胞增殖的抑制作用，衰老相关-β-半乳糖苷酶（SA-β-gal）活性评判细胞衰老，流式细胞术分析细胞周期分布，实时定量PCR法检测78个人类细胞衰老相关基因的表达。结 果 顺铂对CNE2细胞的增殖具有明显抑制作用；1.0 mg/L顺铂处理后第6和第8天检测SA-β-gal染色阳性细胞数分别达到67.63%和89.22%，细胞阻滞于G2 期；TP53、TP63、p16Ink4a，p27Kip1、PTEN、RB1、Cdc25C、TBX3、Gadd45a、IGF1R、PIK3CA、BMI-1、B2M、MORC3、MYC和SPARC等16个衰老相关基因的mRNA水平显著升高（P＜0.05或0.01），Cyclin A2、Cyclin B1、Cyclin E1、CDK6、ATM、ETS2、MDM2、E2F1、ETS1、FN1、IGFBP3、RBL2、SERPINB2及SIRT1等14个基因表达水平下降（P＜0.05 或 0.01）。结论 顺铂能够在体外诱导CNE2细胞衰老，其机制涉及p16和p27介导的p53-pRB和PTEN衰老信号调控网络。

Cisplatin-induced Senescence and Senescence-related Gene Expression Profiles in#br# Human Nasopharyngeal Carcinoma Cells CNE2

Objective To investigate the effects of cisplatin induction in vitro on human nasopharyngeal carcinoma cells CNE2 and its possible molecular mechanism. Methods Proliferation of CNE2 cells treated with various concentrations(0.4-4.0 mg/L) of cisplatin was determined using MTT assay. The senescent cells were determined by a senescence-associated-β-galactosidase(SA-β-gal) activity assay and cell cycle distribution was analyzed by flow cytometry. A Sybe Green real-time polymerase chain reaction(RT-PCR) method was used to examine the changes of expression profiles of 78 human cellular senescence-related genes. Results Cisplatin inhibited the proliferation of CNE2 cells in a dose- and time-dependent manner, and significantly induced a cellular senescence in CNE2 cells, which respectively counted at 67.63% of senescent cells at day 6 and 89.22% at day 8, after treatment with 1.0 mg/L of cisplatin. A G2/M phase arrest was observed. The expression levels of 16 senescence-related genes including TP53, TP63, p16Ink4a, p27Kip1, PTEN, RB1, TBX3, Cdc25C, Gadd45a, IGF1R, PIK3CA, MI-1, B2M, MORC3, MYC and SPARC were increased(P＜0.05 or 0.01), while the 14 genes including Cyclin A2, Cyclin B1, Cyclin E1,CDK6, ATM, E2F1, ETS1, ETS2, MDM2, FN1, IGFBP3, RBL2, SERPINB2 and SIRT1 were decreased at mRNA levels(P＜0.05 or 0.01). Conclusion Cisplatin could induce G2/M phase arrest and cellular senescence in CNE2 cells in vitro, which may involve a omplicated network of p53-pRb signaling or/and PTEN signaling pathways mediated by p16 and p27.