芹菜素对胰腺癌BxPC-3细胞Sonic Hedgehog信号通路调控和自我更新能力的影响  

Effect of Apigenin on Sonic Hedgehog Signaling Pathway Regulation and SelfRenewal Ability in Pancreatic Cancer BxPC-3 Cells

作　　者：潘岩 刘鲁明 花永强 楼银妹 PAN Yan;LIU Luming;HUA Yongqiang;LOU Yinmei

机构地区：[1]杭州市富阳区第一人民医院中西医结合科,浙江杭州311400 [2]复旦大学附属肿瘤医院中西医结合科,上海200032

出　　处：《新中医》2022年第9期134-138,共5页New Chinese Medicine

基　　金：杭州市科技计划引导项目(20171226Y162)。

摘　　要：目的:探讨芹菜素对胰腺癌 BxPC-3 细胞 Sonic Hedgehog 信号通路的调控作用和自我更新能力的影响。方法:用不同浓度 (10、20 和 50 μmol/L) 芹菜素干预胰腺癌 BxPC-3 细胞。应用流式细胞仪法检测胰腺癌肿瘤干细胞 (CD24^(+)CD44^(+)) 比例,利用 RT-PCR 和 Western blot 法检测 Sonic Hedgehog 信号通路成员 SHH、PTCH1、SMO 和 GLI1 在 mRNA 和蛋白水平的表达情况,采用无血清培养成球实验观察胰腺癌细胞自我更新能力。结果:BxPC-3 细胞经不同浓度 (10、20 和 50 μmol/L) 芹菜素干预 48 h 后,胰腺癌 CD24^(+)CD44^(+)细胞比例均呈不同程度的下降 (P<0.01)。与对照组比较,20、50 μmol/L 芹菜素可显著下调胰腺癌肿瘤干细胞比例 (P<0.01),且显著抑制 Hedgehog 信号通路成员 SHH、PTCH1、SMO 和 GLI1 在 mRNA 和蛋白水平的表达 (P<0.01)。无血清培养的 BxPC-3 细胞经不同浓度 (20 和 50 μmol/L) 芹菜素干预 7 d 后,肿瘤干细胞成球数目较对照组显著减少 (P<0.01)。结论:芹菜素抑制胰腺癌细胞的自我更新能力与下调 Sonic Hedgehog 信号通路有关。Objective:To investigate the effect of apigenin on regulation of Sonic Hedgehog signaling pathway and selfrenewal capacity of pancreatic cancer BxPC-3 cells.Methods:Different concentrations of apigenin(10,20,and 50μmol/L)were used to intervene in pancreatic cancer cells.The proportion of the pancreatic cancer stem cell,CD24~+CD44~+,was detected by the method of flow cytometry.The expressions of SHH,PTCH1,SMO and GLI1 in the Sonic Hedgehog signaling pathway at mRNA and protein levels were detected by the method of RT-PCR and Western blot.The self-renewal ability of pancreatic cancer cells was observed by the sphere formation assay with serum-free culture.Results:After BxPC-3cells were intervened with different concentrations of apigenin(10,20 and 50 μmol/L) for 48 hours,the proportion of CD24~+ CD44~+,the pancreatic cancer cell,was decreased in varying degrees(P<0.01).Compared with the control group,20 and 50 μmol/L of apigenin significantly decreased the proportion of pancreatic cancer stem cells(P<0.01),and significantly inhibited the expression of SHH,PTCH1,SMO and GLI1 at mRNA and protein levels(P<0.01).After the BxPC-3 cells in serum-free medium were treated with different concentrations of apigenin(20 and 50 μmol/L) for seven days,the number of tumor stem cell spheres was significantly reduced when compared with that in the control group(P<0.01).Conclusion:Apigenin inhibiting the self-renewal of pancreatic cancer cells is related to the down-regulation of Sonic Hedgehog signaling pathway.

关 键 词：胰腺癌 BXPC-3细胞 芹菜素 Sonic Hedgehog 自我更新 GLI1 

分 类 号：R735.9[医药卫生—肿瘤]