【目的】 探讨蟾毒灵调节LIM激酶1（LIMK1）-Cofilin1通路对肺癌细胞增殖、迁移和上皮间质转化的影响。【方法】 培养 肺癌细胞A549，将其分成对照组、蟾毒灵组（40 ng/mL）、LIMK1组（转染pcDNA-LIMK1质粒）、蟾毒灵+NC组（40 ng/mL蟾毒 灵+转染pcDNA-NC质粒）及蟾毒灵+LIMK1组（40 ng/mL蟾毒灵+转染pcDNA-LIMK1质粒）。克隆形成实验观察细胞克隆形成 情况；MTT法检测细胞活性；划痕愈合实验检测细胞迁移能力；Transwell实验检测细胞侵袭能力；流式细胞术检测细胞凋 亡水平；Western Blot法检测细胞凋亡、上皮间质转化和 LIMK1-Cofilin1通路相关蛋白表达水平。【结果】 与对照组比较，蟾 毒灵组细胞克隆形成率、MTT 吸光度（OD）值、划痕愈合率、细胞侵袭数及 B 细胞淋巴瘤/白血病 2（Bcl-2）、波形蛋白 （Vimentin）、N-钙黏着蛋白（N-cadherin）、磷酸化（p）-LIMK1和p-Cofilin1蛋白表达量降低（P＜0.05），细胞凋亡率及Bcl-2相 关 X 蛋白（Bax）、上皮钙黏蛋白（E-cadherin）蛋白表达量增加（P＜0.05），而LIMK1组细胞克隆形成率、MTT OD值、划痕愈合 率、细胞侵袭数及 Bcl-2、Vimentin、N-cadherin、p-LIMK1 和 p-Cofilin1 蛋白表达量增加（P＜0.05），细胞凋亡率及 Bax、 E-cadherin蛋白表达量下降（P＜0.05）；与蟾毒灵+NC组比较，蟾毒灵+LIMK1 组细胞克隆形成率、MTT OD 值、划痕愈合率、 细胞侵袭数及 Bcl-2、Vimentin、N-cadherin、p-LIMK1 和 p-Cofilin1 蛋白表达量增加（P＜0.05），细胞凋亡率及 Bax、 E-cadherin 蛋白表达量减少（P＜0.05）。与LIMK1组比较，蟾毒灵+LIMK1组克隆形成率、MTT OD值、划痕愈合率、细胞侵袭数 及Bcl-2、Vimentin、N-cadherin、p-LIMK1和p-Cofilin1蛋白表达量降低（P＜0.05），细胞凋亡率及Bax、E-cadherin蛋白表达 量增加（P＜0.05）。【结论】 蟾毒灵可能通过抑制LIMK1-Cofilin1通路抑制肺癌细胞增殖、迁移和上皮间质转化。

Objective To investigate the effects of bufalin on the proliferation， migration， and epithelialmesenchymal transition （EMT） of lung cancer cells via regulation of the LIM kinase 1（LIMK1）-Cofilin1 pathway. Methods Lung cancer A549 cells were cultured and divided into the following groups：control，bufalin （40 ng/mL）， LIMK1（transfected with pcDNA-LIMK1 plasmid），bufalin+NC （40 ng/mL bufalin + pcDNA-NC plasmid），and bufalin+LIMK1 （40 ng/mL bufalin + pcDNA-LIMK1 plasmid）. Colony formation assay was used to observe clonogenicity；MTT assay was used to determine cell viability；scratch wound healing assay was used to evaluate cell migration；Transwell assay was used to measure cell invasion；flow cytometry was used to detect apoptosis；and Western Blot was used to examine the expression of proteins related to apoptosis，EMT，and the LIMK1- Cofilin1 pathway. Results Compared with the control group，the bufalin group showed decreased colony formation rate，MTT optical density （OD） value，scratch healing rate，number of invading cells，and protein expression levels of B-cell lymphoma/leukemia-2（Bcl-2），vimentin，N-cadherin，phosphorylated （p）-LIMK1，and pCofilin1（P＜0.05），while apoptosis rate and protein expression levels of Bcl-2-associated X protein （Bax） and E-cadherin were increased （P＜0.05）. In contrast，the LIMK1 group exhibited increased colony formation rate， MTT OD value，scratch healing rate，number of invading cells，and protein expression levels of Bcl-2，vimentin， N-cadherin， p-LIMK1， and p-Cofilin1（P＜0.05）， while apoptosis rate and expression levels of Bax and E-cadherin were decreased （P＜0.05）. Compared with the bufalin+NC group， the bufalin+LIMK1 group demonstrated increased colony formation rate （P＜0.05），MTT OD value，scratch healing rate，number of invading cells，and protein expression levels of Bcl-2，vimentin，N-cadherin，p-LIMK1，and p-Cofilin1（P＜0.05）， while apoptosis rate and expression levels of Bax and E-cadherin were decreased （P＜0.05）. Compared with the LIMK1 group，the bufalin+LIMK1 group showed reduced colony formation rate，MTT OD value，scratch healing rate，number of invading cells，and protein expression levels of Bcl-2，vimentin，N-cadherin，p-LIMK1，and p-Cofilin1（P＜0.05），while apoptosis rate and protein expression levels of Bax and E-cadherin were increased （P＜0.05）. Conclusion Bufalin may inhibit the proliferation， migration， and EMT of lung cancer cells by suppressing the LIMK1-Cofilin1 pathway.

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陕西省教育厅专项科研计划项目（编号：20JK0600）；陕西中医药大学学科建设基金项目（编号：2020XK05）