炎性因子对人胰腺癌PaTu8988细胞NF-KB及Hedgehog通路成员表达的影响

Pro-inflammatory cytokines inhibit apoptosis of pancreatic ductal adenocarcinoma cells via both NF-kB and Hedgehog signaling pathways

目的探讨肿瘤坏死因子仅（TNF-α）和白介素lp（IL-1β）对人胰腺癌PaTu8988细胞核因子KB（NF-KB）及Hedgehog（HH）通路成员Shh、SMO、Glil、SuFu基因表达的影响。方法分别应用TNF-α和IL-1β刺激人胰腺癌PaTu8988细胞48h，以未处理细胞作为对照组。采用实时定量RT-PER和蛋白质印迹法检测各组细胞NF-KB及Shh、SMO、Glil、SuFu的mRNA和蛋白表达，采用流式细胞仪检测各组细胞的凋亡。结果TNF-α刺激组、IL-18刺激组、对照组PaTu8988细胞的NF-KBmRNA表达量分另0为9．92±0．78、7．74±0．32、1．01±0．08；GlilmRNA为7．25±0．45、5．74±0．33、1．00±0．06；ShhmRNA为3．60±0．36、4．33±0．45、1．00±0．04；SMOmRNA为1．03±0．15、1．07±0．16、1．01±0．06；SuFumRNA为0．88±0．14、0．96±0．13、1．01±0．05。其中TNF-α刺激组、IL-1β刺激组的NF-KB、Glil、ShhmRNA表达量均较对照组显著增加，差异有统计学意义（P值〈0．05或〈0．01）。两刺激组的NF-KB、Glil、Shh蛋白表达量也较对照组增高，差异具有统计学意义（P值均〈0．05）。TNF-α刺激组、IL-1β刺激组、对照组PaTu8988细胞凋亡率分别为（17．40±2．87）％、（11．05±1．34）％、（49．90±2．96）％，TNF-α刺激组、IL-1β刺激组的细胞凋亡率均较对照组显著减少，差异有统计学意义（P值均〈0．01）。结论TNF-α和IL-1β可激活PaTu8988细胞的NF-KB和HH信号通路的Shh、Glil基因的表达，减少细胞的凋亡。

Objective To investigate the effect of TNF-α and IL-1β on the expression of human pancreatic cancer PaTu8988 NF-KB and Hedgehog （HH） signaling pathways members Glil, Shh, SMO, SuFu. Methods Pancreatic cancer cell line PaTu8988 was treated with TNF-α and IL-1β, respectively, for 48 h, and cells without treatment were control group. Then the protein and mRNA expression of NF-KB and Shh, SMO, Glil, SuFu was determined by RT-PCR and Western blot; apoptosis of cells were tested by flow cytometry. Results The expressions of NF-KB mRNA in PaTu8988 cells in TNF-α group, IL-1βgroup, control group were 9.92±0.78, 7.74±0.32, 1.01±0.08; and the expressions of Gill mRNA were 7.25 ± 0.45, 5.74±0.33, 1.00±0.06; and the expressions of Shh mRNA were 3.60±0.36, 4.33±0.45, 1.00±0.04; and the expressions of SMO mRNA were 1.03 ± 0.15, 1.07 ± 0.16, 1.01 ± 0.06; and the expressions of SuFu mRNA were 0.88 ±0.14, 0.96 ±0.13, 1.01 ±0.05. The expressions of NF-KB, Glil,Shh mRNA in TNF-α group, IL-1β group were significantly higher than that in control group, and the difference between the two groups was statistically significant （ P 〈 0.05 ）. The expressions of corresponding proteins were consistent with the expressions of mRNA. The apoptosis rates in TNF- α group, IL-1β group, control group were （ 17.40 ± 2.87 ） %, （ 11.05 ± 1.34） %, （49.90 ± 2.96） %, and the apoptosis rates in TNF-α group, IL-1β group were significantly lower than that in control group, and the difference between the two groups was statistically significant （P 〈0. 01 ）. Conclusions TNF-α and IL-1βα can activate the expression of NF-KB and Shh, Glil in human pancreatic cancer PaTu898, and decrease apoptosis.