TLR9激动剂在胰腺星状细胞介导的胰腺癌耐药中的作用

目的:研究Toll样受体9(Toll-like receptor 9,TLR9)激动剂在胰腺星状细胞(pancreatic stellate cells,PSCs)介导的胰腺癌化疗耐药中的作用。方法:分离并培养小鼠胰腺星状细胞,采用免疫组化检测小鼠胰腺星状细胞TLR9的表达情况;应用MTT法检测TLR9激动剂OND1826对胰腺星状细胞增殖的影响;将胰腺星状细胞与胰腺癌MIAPaCa-2细胞进行Transwell共培养,分为对照组,共培养组,共培养+OND1826组,48 h后采用ELISA法检测上清液中的氧化亚氮(NO)及白介素-1β(IL-1β)表达量,加入10μg/ml吉西他滨继续培养24 h,采用MTT法检测肿瘤细胞凋亡情况。结果:免疫组化结果显示胰腺星状细胞表达TLR9,TLR9激动剂可抑制胰腺星状细胞生长(P<0.05),与对照组相比,共培养组的NO、IL-1β表达量明显增加,肿瘤细胞凋亡明显减少(P<0.05);OND1826干预后NO、IL-1β表达量有所减少,而肿瘤细胞凋亡有所增加(P<0.05)。结论:TLR9激动剂可抑制胰腺星状细胞的增殖能力,进而抑制星状细胞介导的胰腺癌耐药。

Significance of TLR9 receptor agonists in pancreatic stellate cell mediated pancreatic cancer resistance

Objective: To research role of Toll-like receptor 9(TLR9) agonist in pancreatic stellate cells(PSCs) and it mediated pancreatic cancer drug resistance.Methods: Isolating and culturing mouce PSCs,and using immunohistochemistry to demonstrate its TLR9 expression.Using MTT to assay PSCs proliferation by TLR9 agonist.PSCs and MIAPaCa-2 cells were co-cultured in Transwell units,which were divided into control group,co-cultured group,co-culture+OND1826 group.ELISA was used to detect nitric oxide(NO) and interleukin-1β(IL-1β) in supernatant after 48 hours.MIAPaCa-2 cells were cultured in another 24 hours by adding 10 μg/ml of gemcitabine and detected tumor cell apoptosis rate by MTT.Results: Immunohistochemistry showed that PSCs expressed TLR9.Proliferation of PSCs was inhibited by using TLR-9 agonists(P<0.05).Compared with the control group,co-cultured group had a higher expression level of NO,IL-1β,and a lower apoptosis rate(P<0.05).NO,IL-1β expression levels decreased,and apoptosis rate increased by OND1826(P<0.05).Conclusion: PSCs mediated pancreatic cancer drug resistance could be inhibited by TLR9 agonists.