目的 探討X連鎖凋亡抑制蛋白（XIAP）抑制劑蒽貝素抑制人胰腺癌細(xì)胞Mia PaCa-2凋亡的作用及其可能機(jī)制。方法 MTT法和流式細(xì)胞術(shù)檢測(cè)不同劑量蒽貝素對(duì)Mia PaCa-2細(xì)胞增殖和凋亡的影響；RT-PCR法對(duì)蒽貝素處理前后Mia PaCa-2細(xì)胞中XIAP基因的表達(dá)進(jìn)行檢測(cè)。Western blotting方法檢測(cè)蒽貝素作用后細(xì)胞XIAP、Caspase 3、促凋亡蛋白Bax和抗凋亡蛋白Bcl-2表達(dá)的變化。結(jié)果 蒽貝素對(duì)人胰腺癌細(xì)胞增殖具有顯著抑制作用（P<0.05），經(jīng)不同劑量蒽貝素處理后Mia PaCa-2細(xì)胞凋亡率明顯增高，與未處理組比較差異具有顯著性（P<0.01）；RT-PCR顯示對(duì)照組細(xì)胞中XIAP的-△△CT值是實(shí)驗(yàn)組的11.31倍；經(jīng)蒽貝素作用24 h后，Mia PaCa-2細(xì)胞出現(xiàn)Caspase 3裂解片段，且Bax/Bcl-2比值增大。結(jié)論 蒽貝素在體外可顯著抑制Mia PaCa-2細(xì)胞增殖，誘導(dǎo)細(xì)胞凋亡，其機(jī)制可能是通過(guò)拮抗XIAP的作用，激活Caspase相關(guān)性的細(xì)胞凋亡內(nèi)源性途徑而誘導(dǎo)人胰腺癌細(xì)胞發(fā)生凋亡。

Objective To discuss the effect of X-linked inhibitor of apoptosis (XIAP) inhibitor embelin on the apoptosis of pancreatic cancer Mia PaCa-2 cells and its possible mechanism. Methods The growth suppression and the apoptotic rate of embelin on Mia PaCa-2 cells were evaluated by MTT assay and flow cytometry. Real-time quantitative reverse transcriptionpolymerase chain reaction (RT-PCR) method was used to detect the expression of XIAP gene in Mia PaCa-2 cells with or without treatment of embelin. Western blotting analysis was performed to assess the expression of XIAP, Caspase 3, pro-apoptotic factor Bax as well as the pro-survival factor Bcl-2. Results Embelin inhibited the proliferation of the pancreatic cancer cells (P<0.05). The apoptosis of Mia PaCa-2 cells were significantly increased after embelin treatment at different concentration as compared with that in the control group (P<0.01). Furthermore, the -△△CT value of XIAP in the control group was 11.31 times as much as that in the treatment group. The cleavage of Caspase 3 was observed after 24 h treated with embelin. Moreover, Bax expression was upregulated while Bcl-2 levels were decreased. Conclusion Embelin can significantly inhibit the proliferation of Mia PaCa-2 cells, induce the apoptosis of Mia PaCa-2 cells by antagonizing the endogenous XIAP pathway that activates caspase-dependent apoptosis in Mia PaCa-2 cells.

天津市衛(wèi)生局重點(diǎn)課題資助項(xiàng)目（2012KR06）