肿瘤坏死因子相关凋亡诱导配体（TRAIL）和牛磺罗定可诱导人纤维肉瘤细胞凋亡并抑制其增殖。

TRAIL and Taurolidine induce apoptosis and decrease proliferation in human fibrosarcoma.

作者信息

Daigeler Adrien, Brenzel Christina, Bulut Daniel, Geisler Anne, Hilgert Christoph, Lehnhardt Marcus, Steinau Hans U, Flier Annegret, Steinstraesser Lars, Klein-Hitpass Ludger, Mittelkötter Ulrich, Uhl Waldemar, Chromik Ansgar M

机构信息

Department of Plastic Surgery, Burn Center, Hand Center, Sarcoma Reference Center, BG-University Hospital Bergmannsheil, Bürkle-de-la-Camp-Platz 1, 44789 Bochum, Germany.

出版信息

J Exp Clin Cancer Res. 2008 Dec 12;27(1):82. doi: 10.1186/1756-9966-27-82.

BACKGROUND

Disseminated soft tissue sarcoma still represents a therapeutic dilemma because effective cytostatics are missing. Therefore we tested TRAIL and Tarolidine (TRD), two substances with apoptogenic properties on human fibrosarcoma (HT1080).

METHODS

Viability, apoptosis and necrosis were visualized by TUNEL-Assay and quantitated by FACS analysis (Propidiumiodide/AnnexinV staining). Gene expression was analysed by RNA-Microarray and the results validated for selected genes by rtPCR. Protein level changes were documented by Western Blot analysis. NFKB activity was analysed by ELISA and proliferation assays (BrdU) were performed.

RESULTS AND DISCUSSION

The single substances TRAIL and TRD induced apoptotic cell death and decreased proliferation in HT1080 cells significantly. Gene expression of several genes related to apoptotic pathways (TRAIL: ARHGDIA, NFKBIA, TNFAIP3; TRD: HSPA1A/B, NFKBIA, GADD45A, SGK, JUN, MAP3K14) was changed. The combination of TRD and TRAIL significantly increased apoptotic cell death compared to the single substances and lead to expression changes in a variety of genes (HSPA1A/B, NFKBIA, PPP1R15A, GADD45A, AXL, SGK, DUSP1, JUN, IRF1, MYC, BAG5, BIRC3). NFKB activity assay revealed an antipodal regulation of the several subunits of NFKB by TRD and TRD+TRAIL compared to TRAIL alone.

CONCLUSION

TRD and TRAIL are effective to induce apoptosis and decrease proliferation in human fibrosarcoma. A variety of genes seems to be involved, pointing to the NFKB pathway as key regulator in TRD/TRAIL-mediated apoptosis.

背景

由于缺乏有效的细胞抑制剂，播散性软组织肉瘤仍然是一个治疗难题。因此，我们测试了TRAIL和塔里多定（TRD）这两种对人纤维肉瘤（HT1080）具有凋亡诱导特性的物质。

方法

通过TUNEL检测可视化细胞活力、凋亡和坏死情况，并通过流式细胞术分析（碘化丙啶/膜联蛋白V染色）进行定量。通过RNA微阵列分析基因表达，并通过逆转录聚合酶链反应对选定基因的结果进行验证。通过蛋白质印迹分析记录蛋白质水平变化。通过酶联免疫吸附测定分析核因子κB活性，并进行增殖测定（5-溴-2'-脱氧尿苷）。

结果与讨论

单一物质TRAIL和TRD可显著诱导HT1080细胞凋亡并降低其增殖。与凋亡途径相关的几个基因的表达发生了变化（TRAIL：ARHGDIA、NFKBIA、TNFAIP3；TRD：HSPA1A/B,、NFKBIA、GADD45A、SGK、JUN、MAP3K14）。与单一物质相比，TRD和TRAIL的组合显著增加了凋亡细胞死亡，并导致多种基因的表达变化（HSPA1A/B、NFKBIA、PPP1R15A、GADD45A、AXL、SGK、DUSP1、JUN、IRF1、MYC、BAG5、BIRC3）。核因子κB活性测定显示，与单独的TRAIL相比，TRD和TRD+TRAIL对核因子κB的几个亚基具有相反的调节作用。