穿心莲内酯调节 SHH/GLI1 信号通路对直肠癌细胞活性和化疗耐药性的影响

Effect of andrographolide on the activity and chemotherapy resistance of rectal cancer cells by regulating the SHH/GLI1 signaling pathway

关键词:  直肠癌  穿心莲内酯  SHH/GLI1信号通路  化疗耐药性

Key words:  rectal cancer  andrographolide  SHH/GLI1 signaling pathway  chemotherapy resistance

摘要:

[目的] 探讨穿心莲内酯调节声波刺猬（SHH）/GLI家族锌指蛋白（GLI1）信号通路对直肠癌细胞活性和化疗耐药性的影响。[方法] 采用3.75~120 μmol/L穿心莲内酯处理人结直肠癌耐药细胞株SW20/5-FU，检测细胞增殖活力，筛选穿心莲内酯实验浓度。将SW20/5-FU细胞随机分为对照组（Control组）、穿心莲内酯低、中、高浓度组（AND-L组、AND-M组、AND-H组）、高浓度穿心莲内酯+SHH信号通路抑制剂组（Cyclopamine组）；检测SW20/5-FU细胞的增殖、迁移、侵袭能力；检测上皮间质转化（EMT）和通路蛋白表达。[结果] 通过检测细胞增殖活力，选用15.0、30.0、60.0 μmol/L的穿心莲内酯作处理浓度。与Control组比较，AND-L、AND-M、AND-H组细胞克隆形成数、划痕愈合率、侵袭数量降低，细胞周期蛋白D1（CyclinD1）、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）、神经钙黏蛋白（N-Cadherin）、波形蛋白（Vimentin）表达水平下降，上皮性黏附蛋白（E-Cadherin）、SHH、GLI1蛋白表达水平升高（P<0.05）；与AND-H组比较，Cyclopamine组克隆形成数、划痕愈合率、侵袭数量上升，CyclinD1、MMP-2、MMP-9、N-Cadherin、Vimentin表达水平升高，E-Cadherin、SHH、GLI1蛋白表达水平降低（P<0.05）。[结论] 穿心莲内酯激活SHH/GLI1信号通路抑制直肠癌细胞增殖、迁移、侵袭与EMT能力，从而改善化疗耐药性。

Abstract:

[Objective] To investigate the effect of andrographolide on the activity and chemotherapy resistance of rectal cancer cells by regulating the sonic hedgehog（SHH）/GLI family zinc finger protein 1（GLI1） signaling pathway. [Methods] Human colorectal cancer drug-resistant cell line SW20/5-FU was treated with 3.75 to 120 μmol/L of andrographolide，and cell proliferation activity was detected. The experimental concentration of andrographolide was screened. SW20/5-FU cells were randomly divided into Control group，low，medium，and high concentration andrographolide groups（AND-L group，AND-M group，AND-H group），and high concentration andrographolide + SHH signaling pathway inhibitor group（Cyclopamine group）. The proliferation，migration，and invasion abilities of SW20/5-FU cells were detected. Epithelial mesenchymal transition（EMT） and pathway protein expression were detected. [Results] By detecting cell proliferation activity，the concentrations of andrographolide at 15.0，30.0，and 60.0 μmol/L were selected as the treatment concentrations. Compared with the Control group，the number of cell clones formed，scratch healing rate，and number of invasions decreased in AND-L，AND-M，and AND-H groups，while the expression levels of G1/S-specific cyclin D1（Cyclin D1），matrix metalloproteinase-2（MMP-2），matrix metalloproteinase-9（MMP-9），neural cadherin（N-Cadherin），and Vimentin decreased，the expression levels of epithelial cadherin（E-Cadherin），SHH，and GLI1 proteins increased（P<0.05）. Compared with the AND-H group，the number of cell clones formed，scratch healing rate，and number of invasions increased in Cyclopamine group，while the expression levels of CyclinD1，MMP-2，MMP-9，N-Cadherin，and Vimentin increased，the expression levels of E-Cadherin，SHH，and GLI1 proteins decreased（P<0.05）. [Conclusion] Andrographolide activates the SHH/GLI1 signaling pathway to inhibit the proliferation，migration，invasion，and EMT ability of rectal cancer cells，thereby improving chemotherapy resistance.