首页|基于Wnt信号通路的熊果酸调节结直肠癌SW480细胞自噬的研究

基于Wnt信号通路的熊果酸调节结直肠癌SW480细胞自噬的研究

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  • NETL
  • NSTL
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目的:研究熊果酸对结直肠癌细胞自噬的影响及其基于Wnt信号通路的调节自噬机制.方法:MTT法检测熊果酸对SW480细胞增殖的影响;细胞划痕试验检测熊果酸5.47、10.94、21.88 μmol/L对SW480细胞迁移能力的影响;MDC/PI法检测其对SW480细胞自噬体形成的影响;qRT-PCR法检测对细胞自噬相关基因微观相关蛋白轻链3(Lc3)、苄氯素1(Beclin1)、自噬相关基因5(Atg5)和自噬蛋白(P62)以及Wnt信号通路相关基因β连环蛋白(Bcatenin)、T-细胞因子4(Tcf4)、糖原合成酶激酶3β(Gsk3β)、Wnt家族成员5A(Wnt5a)和Myc原癌基因(Cmyc)mRNA表达的影响;Western Blot法检测熊果酸对细胞自噬相关蛋白LC3-Ⅱ、BEC-LIN-1、ATG5 和 P62 以及 Wnt 信号通路相关蛋白 β-catenin、NUCLEAR β-catenin、TCF4、GSK-3β、p-GSK-3β、WNT5A 和 C-MYC 表达的影响.结果:熊果酸能显著抑制SW480细胞增殖和迁移(P＜0.01);增强SW480细胞自噬体荧光强度(P＜0.01);熊果酸能显著上调SW480 细胞Lc3、Beclin1、Atg5 和 Gsk3b mRNA 表达,下调 P62、Bcatenin、Tcf4、Wnt5a 和 Cmyc mRNA 表达(P＜0.05),显著上调 SW480细胞 LC3-Ⅱ、BECLIN-1、ATG5 和 GSK-3β 蛋白表达,下调 P62、β-catenin、NUCLEAR β-catenin、TCF4、WNT5A、p-GSK-3β(Ser389)和 C-MYC蛋白表达(P＜0.05);熊果酸抗SW480细胞增殖作用可被Wnt信号通路抑制剂IWR-1协同增强(P＜0.01),而被Wnt信号通路激动剂SKL2001拮抗减弱(P＜0.01).结论:熊果酸能够抑制SW480细胞增殖、迁移,增强SW480细胞自噬,其机制与降低结直肠癌细胞Wnt信号通路活性相关.
Study on Regulation of Autophagy in Colorectal Cancer SW480 Cells by Ursolic Acid Based on Wnt Signaling Pathway
Objective:To investigate the effect of ursolic acid(UA)on autophagy in colorectal cancer cells and its mechanism of regulation of auto-phagy based on Wnt signaling pathway.Methods:The effect of UA on the proliferation of SW480 cells was detected by MTT assay.In vitro scratch assay was used to measure the effect of UA(5.47,10.94 and 21.88 μmol/L)on the migration of SW480 cells.The influence of UA on autophagosome formation in SW480 cells was observed with MDC/PI assay.The effects of UA on the mRNA levels of autophagy-related genes including microtubule-associated protein light chain3(Lc3),Beclin-1,autophagy-related gene5(Atg5)and P62,and Wnt signaling pathway-related genes involving β-catenin,Tcf4,Gsk-3β,Wnt5a and C-myc were detected by qRT-PCR.Western blot was employed to analyze the influence of UA on the expression of autophagy-related proteins including LC3-Ⅱ,Beclin-1,ATG5 and P62,and Wnt signaling pathway-re-lated proteins involving β-catenin,nuclear β-catenin,TCF4,GSK-3β,p-GSK-3β,Wnt5 A and C-Myc.Results:U A significantly inhibited the proliferation and migration of SW480 cells(P＜0.01),and enhanced the fluorescence intensity of autophagosomes in SW480 cells(P＜0.01).UA significantly up-regulated the mRNA expression levels of Lc3,Beclin-1,Atg5 and Gsk-3β,while down-regulated the mRNA levels of P62,β-catenin,Tcf4,Wnt5a and C-myc in SW480 cells(P＜0.05).Also,UA significantly up-regulated the protein expressions of LC3-Ⅱ,Bec-lin-1,ATG5 and GSK-3β,while down-regulated the protein expressions of P62,β-catenin,nuclear β-catenin,TCF4,Wnt5A,p-GSK-3β(Ser389)and C-Myc in SW480 cells(P＜0.05).The inhibition of UA on the proliferation of SW480 cells was synergized by Wnt signaling in-hibitor IWR-1(P＜0.01),but antagonized by Wnt signaling activator SKL2001(P＜0.01).Conclusion:UA may inhibit the proliferation and migration,and enhance the autophagy of SW480 cells.Its mechanism may be related to decreasing Wnt signaling pathway activity in colorectal cancer cells.

Ursolic acidWnt signaling pathwayAutophagyColorectal cancer

陶秋、王敏敏、雷家荣、彭宇辉、梁园菁、明天琪、唐顺、周丽娟、徐海波

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西南特色中药资源国家重点实验室,成都中医药大学药学院药理系,成都 611137

四川省中医药科学院,成都 610041

熊果酸 Wnt信号通路 自噬 结直肠癌

2024

中药药理与临床
中国药理学会 四川省中医药科学院

中药药理与临床

北大核心
影响因子：0.996
ISSN：1001-859X
年,卷(期)：2024.40(11)
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