摘要
目的研究程序性细胞死亡因子4(PDCD4)诱导的对甲状腺癌SW579细胞凋亡效应及抑制成瘤能力的可能作用机制。方法将对数生长期的细胞分为两组,观察组先予以PDCD4凋亡相关基因Bax的刺激14 h进行细胞的收集,然后加入t PA刺激细胞进行收集。对照组未加PDCD4凋亡相关基因Bax的刺激和t PA刺激。应用Annexin V/PI和API等标记,Cyclins/DNA双标流式细胞仪检测甲状腺癌SW579细胞的凋亡及周期特异性,运用RT-PCR的方法检查Caspase-3、Caspase-9活化情况。结果处于静止期的外周血对PDCD4凋亡诱导不敏感,而G1期PDCD4诱导甲状腺癌SW579细胞出现了明显的细胞凋亡。PDCD4诱导的细胞凋亡主要是在细胞周期的G1期发生。观察组的Caspase-3、Caspase-9 m RNA的表达值分别为(1.12±0.56)、(1.10±0.29),显著高于对照组的(0.82±0.31)、(0.72±0.26),差异有统计学意义(P<0.05)。结论 PDCD4诱导的细胞凋亡与甲状腺癌SW579细胞的细胞周期相关,存在G1期细胞周期阻滞的周期特异性,且此过程中发生了Caspase活化,参与细胞凋亡及可能抑制细胞成瘤能力。
Objective To study the mechanism of programmed cell death 4(PDCD4)-induced apoptosis and inhibition of tumorigenic capacity in SW579 thyroid cancer(TC) cells. Methods SW579 thyroid cancer cells in logarithmic growth phase were divided into two groups. Cells in the observation group were stimulated by PDCD4 apoptosis-related gene Bax for 14 hours and then by t PA for collection. Cells in the control group did not received stimulation by Bax and t PA. Annexin V/PI, API markers and Cyclins/DNA flow cytometry were used to check apoptosis and cycle specificity of SW579 cells. RT-PCR was applied to check the activation conditions of Caspase-3, Caspase-9. Results SW579 cells of peripheral blood in stationary phase were not sensitive to PDCD4 induction, while those in G1 phase showed significant apoptosis under PDCD4 induction. PDCD4-induced apoptosis mainly occurred in the G1 phase. The expression levels of Caspase-3, Caspase-9 m RNA in the observation group were(1.12±0.56),(1.10±0.29), which were significantly higher than those in the control group of(0.82±0.31),(0.72±0.26), P<0.05. Conclusion PDCD4-induced apoptosis of SW579 cells is related to cell cycle, with cycle specificity of cell cycle arrest in G1 phase. During the cell cycle arrest, Caspase activation occurred and is involved in the apoptosis and inhibition of tumorigenic capacity.
出处
《海南医学》
CAS
2016年第23期3790-3793,共4页
Hainan Medical Journal
基金
广东省深圳市福田区公益性科研项目(编号:FTWS2014010)
