Fang Wenli, Yue Shujun, Gan Delu, Zhang Dian, Shi He, Yao Mengli, Qian Husun, Zhou Ting, Chen Tingmei
Ministry-of-Education Key Laboratory of Laboratory Medical Diagnostics, Chongqing Medical University, Chongqing 400016, China.
Ministry-of-Education Key Laboratory of Laboratory Medical Diagnostics, Chongqing Medical University, Chongqing 400016, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2021 Feb;37(2):125-131.
Objective To investigate the effect of progranulin (PGRN) on the invasion and migration of mouse breast cancer 4T1 cells and its mechanism. Methods After treated with PGRN (1 μg/mL) for 24 hours, the invasion ability of breast cancer 4T1 cells was detected by Transwell invasion assay, the migration ability was detected by scratch test, and the epithelial cadherin (E-cadherin), vimentin mRNA expression was detected by real-time fluorescent quantitative PCR. Western blot assay was used to detect the expression of E-cadherin, vimentin, extracellular signal-regulated kinase 1/2 (ERK1/2) and phosphorylated ERK1/2 (p-ERK1/2). After treated with 1 μg/mL PGRN and ERK1/2 signaling pathway inhibitor U0126 (10 μmol/L) simultaneously, the migration and invasion ability of 4T1 cells and the changes in the expression of E-cadherin, vimentin and p-ERK proteins were detected again. Results After treated with PGRN, the migration and invasion capabilities of breast cancer 4T1 cells were significantly enhanced; E-cadherin expression decreased; vimentin and p-ERK1/2 expression increased. After treated with ERK1/2 signaling pathway inhibitor, the ability of PGRN to promote breast cancer 4T1 cell migration, invasion and epithelial-mesenchymal transition (EMT) was significantly inhibited. Conclusion PGRN can promote the migration and invasion of breast cancer 4T1 cells by promoting EMT and activating the ERK1/2 pathway.
目的 探讨颗粒蛋白前体（PGRN）对小鼠乳腺癌4T1细胞侵袭和迁移能力的影响及其机制。方法 用PGRN（1 μg/mL）处理24小时后，采用Transwell侵袭实验检测乳腺癌4T1细胞的侵袭能力，划痕实验检测迁移能力，实时荧光定量PCR检测上皮钙黏蛋白（E-cadherin）、波形蛋白mRNA表达。采用蛋白质免疫印迹法检测E-cadherin、波形蛋白、细胞外信号调节激酶1/2（ERK1/2）和磷酸化ERK1/2（p-ERK1/2）的表达。同时用1 μg/mL PGRN和ERK1/2信号通路抑制剂U0126（10 μmol/L）处理后，再次检测4T1细胞的迁移和侵袭能力以及E-cadherin、波形蛋白和p-ERK蛋白表达的变化。结果 用PGRN处理后，乳腺癌4T1细胞的迁移和侵袭能力显著增强；E-cadherin表达降低；波形蛋白和p-ERK1/2表达增加。用ERK1/2信号通路抑制剂处理后，PGRN促进乳腺癌4T1细胞迁移、侵袭和上皮-间质转化（EMT）的能力显著受到抑制。结论 PGRN可通过促进EMT和激活ERK1/2通路来促进乳腺癌4T1细胞的迁移和侵袭。
