山姜素介导NF-κB信号通路对胰腺癌细胞BXPC-3增殖、侵袭和炎症反应的影响

Effects of alpinetin on proliferation,invasion and inflammation response of pancreatic cancer cells BXPC-3 by mediating NF-κB signaling pathway

目的探讨山姜素介导核因子-κB(NF-κB)信号通路对胰腺癌细胞BXPC-3增殖、侵袭能力和炎症反应的影响。方法将人胰腺癌细胞BXPC-3分为对照组(0μmol/L山姜素)和不同浓度(25、50、100、200、400μmol/L)山姜素处理组,细胞计数试剂盒-8(CCK-8)法、克隆形成实验检测细胞增殖能力,流式细胞术检测细胞周期,Transwell小室法检测细胞侵袭,蛋白免疫印迹法检测相关蛋白表达,酶联免疫吸附(ELISA)法检测炎症因子表达情况。结果CCK-8实验结果显示,与对照组比较,不同浓度(25、50、100、200、400μmol/L)山姜素处理后,胰腺癌细胞BXPC-3的细胞活力显著下降(P<0.05);且山姜素浓度越高、处理时间越长,对BXPC-3细胞活性的抑制作用越明显(P<0.05)。与对照组比较,经50、100、200μmol/L山姜素溶液处理后,细胞的克隆形成率、细胞侵袭能力、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、神经-钙黏素(N-cadherin)、纤维连接蛋白(Fibronectin)、NF-κB亚基p65、基质金属蛋白酶-9(MMP-9)、细胞周期蛋白D1(cyclin D1)蛋白表达水平均明显下降,转化生长因子-β(TGF-β)、IL-10、上皮-钙黏素(E-cadherin)及NF-κB抑制蛋白(IκBα)蛋白表达水平明显上升(P<0.05)。加入激活剂脂多糖(LPS)后,细胞活力、细胞侵袭能力、TNF-α、IL-6、p65、MMP-9、cyclin D1蛋白表达水平明显上升,TGF-β、IL-10、IκBα蛋白表达水平明显下降(P<0.05);而加入NF-κB通路抑制剂Sulfasalazine后上述指标变化相反。结论山姜素可抑制胰腺癌细胞BXPC-3的增殖、侵袭能力和炎症反应,其机制可能与介导NF-κB信号通路有关。

Objective To explore the effects of alpinetin on proliferation,invasion and inflammation response of pancreatic cancer cells BXPC-3 by mediating nuclear factor-κB(NF-κB)signaling pathway.Methods The human pancreatic cancer cells BXPC-3 were divided into control group(0μmol/L alpinetin)and different concentrations(25,50,100,200,400μmol/L)of alpinetin groups.The cell proliferation was detected by cell counting kit-8(CCK-8)and clone formation assay.The cell cycle was detected by flow cytometry.The cells invasion was detected by Transwell chamber assay.The expressions of related proteins were detected by Western blotting.The expressions of inflammatory factors were detected by enzyme-linked immunosorbent assay(ELISA).Results CCK-8 results showed that compared with the control group,the activity of pancreatic cancer cells BXPC-3 was significantly decreased after treatment with different concentrations(25,50,100,200,400μmol/L)of alpinetin,and the difference had statistical significance(P<0.05).The higher the alpinetin concentration and the longer the treatment time,the more significant the inhibitory effects on activity of BXPC-3 cells(P<0.05).Compared with the control group,clone formation rate,cell invasion ability,expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),neuro-cadherin(N-cadherin),fibronectin,NF-κB subunit p65,matrix metalloproteinase-9(MMP-9)and cyclin D1(cyclin D1)were significantly decreased,while levels of transforming growth factor-β(TGF-β),IL-10,epithelial-cadherin(E-cadherin)and NF-κB arrestin(IκBα)were significantly increased after treatment with 50,100,200μmol/L alpinetin solution(P<0.05).After adding the activator lipopolysaccharide(LPS),cell activity,cell invasion ability,levels of TNF-α,IL-6,p65,MMP-9 and cyclin D1 were significantly increased,while levels of TGF-β,IL-10 and IκBαwere significantly decreased(P<0.05).However,the changes of the above indexes were reversed after adding NF-κB pathway inhibitor sulfasalazine.Conclusion Alpinetin can inhibit the proliferation,invasion and inflammation response of pancreatic cancer cells BXPC-3,and the mechanism may be related to mediating NF-κB signaling pathway.