摘要
目的探讨骆驼蓬碱(HAR)对子宫内膜癌(EC)细胞增殖、侵袭及上皮间质转化(EMT)的影响及Yes相关蛋白(YAP)/PDZ结合域的转录共刺激因子(TAZ)信号通路在该过程中的作用机制。方法体外培养人EC细胞系Ishikawa,通过CCK8法检测HAR对Ishikawa细胞的毒性;将Ishikawa细胞分为:NC组、HAR低剂量处理(HAR-L)组、HAR高剂量处理(HAR-H)组、HAR-H+YAP/TAZ通路激活剂(PY-60)组;通过克隆形成实验、Transwell侵袭实验分别检测各组Ishikawa细胞增殖和侵袭能力;qRT-PCR实验检测各组Ishikawa细胞中YAP、TAZ mRNA表达量;Western blot检测各组Ishikawa细胞中YAP、TAZ、上皮钙黏蛋白(E-cadherin)、神经钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)蛋白表达量;构建EC裸鼠移植瘤模型,检测各组肿瘤质量、体积及YAP、TAZ蛋白表达量。结果与NC组相比,HAR-L组和HAR-H组Ishikawa细胞集落形成数量、侵袭细胞数、YAP、TAZ mRNA表达量、YAP、TAZ、N-cadherin、Vimentin蛋白表达量明显下降,E-cadherin蛋白表达量明显升高(P<0.05),且HAR-H组中各指标变化趋势更加明显(P<0.05);与HAR-H组相比,HAR-H+PY-60组Ishikawa细胞集落形成数量、侵袭细胞数、YAP、TAZ mRNA表达量、YAP、TAZ、N-cadherin、Vimentin蛋白表达量明显上升,E-cadherin蛋白表达量明显下降(P<0.05);与Model组相比,HAR组裸鼠肿瘤质量、体积及YAP、TAZ蛋白表达量均明显降低(P<0.05)。结论HAR能够抑制EC细胞增殖、侵袭和EMT,其可能与抑制YAP/TAZ信号通路激活有关。
Objective To discuss the effects of harmaline(HAR)on the proliferation,invasion,and epithelial-mesenchymal transition(EMT)of endometrial cancer(EC)cells,and the mechanism of the Yes-associated protein(YAP)/transcriptional coactivator with a PDZ-binding motif(TAZ)signaling pathway in this process.Methods The human EC cell line Ishikawa was cultured in vitro,and the toxicity of HAR to Ishikawa cells was measured by CCK8 assay.Ishikawa cells were classified into NC group,HAR-L low-dose treatment(HAR-L)group,HAR high-dose treatment(HAR-H)group,and HAR-H+YAP/TAZ pathway activator(PY-60)group.The clone formation experiment and Transwell invasion experiment were used to detect the proliferation and invasion abilities of Ishikawa cells in each group.The qRT-PCR experiment was used to detect the YAP and TAZ mRNA in Ishikawa cells of each group.Western blot was used to detect the YAP,TAZ,E-cadherin,N-cadherin,and Vimentin proteins in Ishikawa cells from each group.The EC nude mouse transplant tumor model was constructed,and the tumor mass,volume,and YAP and TAZ proteins were detected in each group.Results For the NC group,the HAR-L group and HAR-H group showed a clear decrease in Ishikawa cell colony formation,invasive cell count,YAP and TAZ mRNAs,YAP,TAZ,N-cadherin,Vimentin proteins,and a clear increase in E-cadherin protein(P<0.05).Moreover,the changes in various indicators were more pronounced in the HAR-H group(P<0.05).For the HAR-H group,the HAR-H+PY-60 group showed a clear increase in Ishikawa cell colony formation,invasive cell count,YAP and TAZ mRNAs,YAP,TAZ,N-cadherin,Vimentin proteins,and a clear decrease in E-cadherin protein(P<0.05).For the Model group,the HAR group showed a clear decrease in tumor mass,volume,and YAP and TAZ proteins in nude mice(P<0.05).Conclusion HAR can inhibit proliferation,invasion,and EMT of EC cells,which may be related to the inhibition of YAP/TAZ signaling pathway activation.
作者
郑佳一
边雪华
ZHENG Jiayi;BIAN Xuehua(Obstetrics and Gynecology Department of Tangshan Maternal and Children Health Hospital,Tangshan,Hebei,063000,China)
出处
《中国优生与遗传杂志》
2025年第9期1896-1901,共6页
Chinese Journal of Birth Health & Heredity
关键词
骆驼蓬碱
YAP/TAZ信号通路
子宫内膜癌
增殖
侵袭
上皮间质转化
harmaline
YAP/TAZ signaling pathway
endometrial cancer
proliferation
invasion
epithelial-mesenchymal transition
