丹参酮ⅡA对人胰腺癌细胞凋亡的诱导作用及其对SAPK/JNK信号转导通路的影响

Tanshinone IIA induces apoptosis of pancreatic cancer cells via the SAPK/JNK signal pathway

目的:研究丹参酮ⅡA诱导人胰腺癌细胞凋亡和凋亡相关基因表达的JNK信号转导通路,揭示其抗胰腺癌的部分机制.方法:MTT法观察丹参酮ⅡA对人胰腺癌PANC-1细胞的生长抑制作用;8、16、32mg/L丹参酮ⅡA分别作用人胰腺癌PANC-1细胞48h后,免疫荧光染色观察细胞凋亡情况;流式细胞仪法(FCM)检测细胞凋亡;Western blot检测丹参酮ⅡA作用PANC-1细胞后SAPK/JNK信号通路的激活情况,荧光定量PCR检测Survivin基因mRNA的表达水平;并比较阻断JNK信号通路后,丹参酮ⅡA对胰腺癌细胞凋亡Survivin基因mRNA的表达.结果:MTT法测得丹参酮ⅡA对人胰腺癌PA N C-1细胞的生长抑制率均有显著的抑制作用,其作用与剂量和作用时间成正相关;丹参酮ⅡA作用48h后,荧光显微镜下观察到经Hoechst染色的典型凋亡细胞.8、16、32mg/L浓度丹参酮ⅡA作用人胰腺癌细胞后的细胞凋亡率分别为8.83%±1.51%,12.86%±2.70%和21.24%±2.58%,与对照组(0.63%±0.18%)比较,均有显著性差异(均P<0.01);阻断JNK信号通路后,凋亡率明显降低(P<0.01).丹参酮ⅡA作用人胰腺癌细胞1h后JNK信号通路被激活,4h达峰值.16mg/L丹参酮ⅡA作用人胰腺癌细胞48h后,Survivin mRNA的表达明显下降,分别为正常细胞的0.61,0.39,0.10倍;阻断JNK信号通路后,丹参酮ⅡA作用人胰腺癌细胞的Survivin mRNA的表达明显上升.结论:丹参酮ⅡA能诱导人胰腺癌PANC-1细胞株凋亡.通过JNK信号转导通路下调Survivin mRNA的表达可能是其诱导胰腺癌细胞凋亡的重要机制.

AIM：To investigate whether tanshinone IIA （TSIIA）induces apoptosis of human pancreaticcancer cells via the SAPK/JNK signal pathway.METHODS：After treatment with TSIIA,MTT assay was used to observe the cytostatic effect of TSIIA on human pancreatic cancer PANC-1 cells;cell apoptosis was assessed by immunofluorescence and flow cytometry（FCM）;p-JNK expression was assayed by Western blot;and mRNA expression of survivin was detected by quantitative fluorescence PCR.RESULTS：TSIIA inhibited PANC-1 cell growth in a concentration-and time-dependent manner.After PANC-1 cells were treated with 8,16,or 32 mg/L of TSIIA for 48 h,typical morphologic changes of apoptosis were observed by fluorescence microscopy after Hoechst staining.The apoptosis rates of cells treated with 8,16,and 32 mg/L of TSIIA for 48 h were（8.83±1.51）%,（12.86±2.70）%and（21.24±2.58）%,respectively,showing a significant difference among the three groups（P0.01）.After the SAPK/JNK signal pathway was blocked,cell apoptosis rate decreased significantly（P0.01）.p-JNK expression began to increase at 1 h and reached the peak at 4 h after TSIIA treatment.The mRNA expression of the survivin gene decreased obviously after treatment with 16 mg/L TSIIA for 48 h but increased significantly when the SAPK/ JNK signal transduction pathway was blocked.CONCLUSION：TSIIA can induce human pancreatic cancer cell apoptosis.TSIIA exerts antipancreatic cancer effects possibly by downregulating the expression of survivin mRNA via the SAPK/JNK signal transduction pathway.