摘要
目的探讨二十二碳六烯酸（DHA）改变凋亡信号通路，引起前列腺癌细胞凋亡的机制。方法利用RT2 Profiler Array-Apoptosis方法，寻找DHA诱导前列腺癌细胞凋亡的作用靶点，可见10个上调和5个下调基因，然后利用实时定量聚合酶链反应（Real-time PCR）方法，验证和筛选结果。结果与对照组比较，DHA处理24 h后，DU145细胞半胱氨酰天冬氨酸特异性蛋白酶（Caspase）家族中的Caspase-1、Caspase-3和Caspase-9转录水平，分别上调了2.06、4.88和12.10倍；促细胞凋亡的B细胞淋巴瘤/白血病-2相关X蛋白（bax）基因上调了2.93倍，并且bax与B细胞淋巴瘤/白血病-2（bcl-2）比值增加；细胞死亡诱导相关基因CIDEA和DFFA，分别上调了2.34和3.21倍；肿瘤坏死因子相关基因LTA和肿瘤坏死因子（TNF），分别上调了2.04和2.24倍；基因警察TP53的mRNA表达上调了2.97倍。另外，DU145细胞线粒体相关凋亡诱导基因（AIFM1）、蛋白激酶（Akt1）、BH3位点死亡诱导基因BID和BIRC6、X连锁凋亡抑制蛋白（XIAP）转录水平的表达，均发生下调。不同浓度DHA刺激24 h后，在前列腺癌DU145细胞Caspase家族中，Caspase-1、Caspase-3、Caspase-9和Caspase-12表达均明显增加，其中Caspase-3、Caspase-9和Caspase-12的表达，与DHA刺激浓度有剂量依赖性关系；前列腺癌DU145细胞bax、CIDEA、DFFA、TNF和肿瘤蛋白P53（TP53）基因表达均明显增加；其中bax、CIDEA和TNF的表达与DHA刺激浓度有剂量依赖性关系；前列腺癌DU145细胞AIFM1、Akt1、BID、BIRC6和XIAP基因表达均明显下降；其中AIFM1、BID和XIAP的表达与DHA刺激浓度有剂量依赖性关系。
结论DHA通过改变Caspase家族等凋亡信号通路相关基因的表达，诱导前列腺癌细胞发生凋亡。
ObjectiveTo explore how docosahexenoic acid （DHA） changes apoptosis signaling pathways to further lead to apoptosis of prostate cancer cells.MethodsRT2 Profiler PCR Arrays were used to screen the targets of DHA-induced prostate cancer cells apoptosis （10 apoptosis genes were up-regulated and 5 apoptosis genes were down-regulated）. Real-time quantitative polymerase chain reaction （Real-time PCR） was used as reliable tools to validate the results.ResultsAs compared with bovine serum albumin （BSA）, after incubation with DHA for 24 h, the transcriptional level of Caspase family： Caspase-1, Caspase-3 and Caspase-9 in DU145 cells were increased 2.06, 4.88 and 12.10 times respectively; The transcriptional level of pro-apoptotic genes B cell lymphoma/leukemia-2 associated X protein （bax） was increased 2.93 times and B cell lymphoma/leukemia-2 （bcl-2）/bax ratio increased. Cell death inducing related genes CIDEA and DFFA were increased 2.34, 3.21 times respectively; Tumor necrosis factor related genes LTA and tumor necrosis factor （TNF） were increased 2.04 and 2.24 times respectively; The expression of tumor protein P53 （TP53） gene mRNA increased 2.97 times. In addition, as compared with the BSA control group, after incubation with DHA for 24 h, the transcriptional levels of apoptosis inducing factor 1 （AIFM1）, Akt1, BH3 locus death inducing gene （BID）, BIRC6 and X-linked inhibitor of apoptosis protein （XIAP） were decreased. After DHA treatment for 24 h, the mRNA expression of Caspase-1, Caspase-3, Caspase-9 and Caspase-12 was increased respectively. Moreover, the expression of Caspase-3, Caspase-9 and Caspase-12 had a dose-dependent relationship with DHA concentration. The mRNA expression of bax, CIDEA, DFFA, TP53 and TNF normalized with TATA box binding protein （TBP） in 24 h DHA-treated DU145 cells was increased obviously, and the expression of bax, CIDEA and TNF had a dose-dependent relationship with DHA concentration. The mRNA expression of AIFM1, protein kinase B （Ak
出处
《中华实验外科杂志》
CSCD
北大核心
2017年第10期1667-1670,共4页
Chinese Journal of Experimental Surgery
基金
河北省卫生厅2017年度指令性计划项目（20170144）
