肿瘤坏死因子相关凋亡诱导配体联合化疗药物杀伤胰腺癌细胞的研究

Synergistic cytotoxic effects of TRAIL and chemotherapeutic agents on pancreatic carcinoma cells

目的研究肿瘤坏死因子相关凋亡诱导配体以及联合应用亚毒性剂量的化疗药物对胰腺癌的治疗作用。方法半定量 RT-PCR 检测 TRAILR mRNA 在胰腺癌细胞株 Canpan-2中的表达。DNA 琼脂糖凝胶电泳检测细胞凋亡情况。应用不同浓度的 TRAIL 及联合亚毒性剂量的氟尿嘧啶(5-fluorouracil)、吉西他滨(gemcitabin)处理胰腺癌细胞,通过 MTT 法检测细胞毒性作用,流式细胞仪分析细胞凋亡率。结果死亡受体 DR4、DR5及诱骗受体 DcR1、DcR2在胰腺癌细胞株 Canpan-2中均有表达。DNA 琼脂糖凝胶电泳可见到典型的凋亡梯形带。TRAIL100 ng/ml 作用胰腺癌细胞24h 后,细胞杀伤率为(29.5±1.2)%,且其作用存在浓度依赖性;联合应用亚毒性剂量的氟尿嘧啶、吉西化滨能够大大提高TRAIL 的细胞毒活性,杀伤率分别为 TRAIL+氟尿嘧啶(43.7±1.4)%,TRAIL+吉西化滨(49.8±1.2)%,联合用药前后差异有显著性(P<0.01)。结论 TRAIL 受体在胰腺癌细胞普遍表达,并存在受体类型的表达差异;TRAIL 与化疗药物氟尿嘧啶、吉西他滨有协同杀伤胰腺癌细胞的作用。

Objective To investigate the synergistic cytotoxic effects of TRAIL and chemotherapeutic agents on pancreatic carcinoma cells. Methods The expression of TRAILR mRNA was assayed by semiquantitative RT-PCR in pancreatic carcinoma cell line of Canpan-2. DNA ladder assay was employed for the determination of cell apoptosis. After the pancreatic carcinoma cells were ex- posed to different concentrations of TRAIL and subtoxic chemotherapeutic agents, the cytotoxicity was detected by MTT assay and the apoptotic rate determined by flow cytometry. Results The expression of death receptor 4 （DR4）, death receptor 5 （DR5）, decoy receptor 1 （DcR1） and decoy receptor 2 （DcR2） was found in the pancreatic carcinoma cells. There were characteristic ladder bands after Canpa-2 was treated by TRAIL （100ng/ml） for 24 h. After the pancreatic carcinoma cells were exposed to 100ng/ml TRAIL for 24h, the killing rate was （29.5±1.2）% and the chemotherapeutic agents dramatically augmented TRAIL-induced cytotoxic function. There was significant difference in the cytotoxicity between the group of single agent and that of agents in combination with TRAIL. Conclusions TRAILR expression is prevalent in pancreatic carcinoma cells with existence of different receptors. The combination of human TRAIL with chemotherapeutic agents such as 5-Fu and gemcitabine can exert synergistic effects on pancreatic carcinoma.