METHODS: The biological role of LIMK1 in regulating the growth, invasion, and metastasis of cervical cancer was studied in SiHa, CaSki cells and nude mice tumor models. The role of LIMK1 in the growth of cervical cancer was evaluated by HE staining. The role of LIMK1 in the invasion, metastasis, and proliferation of cervical cancer was evaluated by cell scratch, Transwell, and monoclonal experiments. The interaction among LIMK1, ROS, and Src was evaluated by Western blotting. The effects of regulating ROS and p-Src expression on LIMK1 in the migration/invasion and proliferation of cervical cancer cells were evaluated through cellular functional assays.
RESULTS: Overexpression of LIMK1 promoted tumor growth in nude mice. Cell scratch, Transwell, and monoclonal experiments suggested that LIMK1 promoted the invasion, metastasis, and proliferation of cervical cancer cells. Western blotting suggested that LIMK1 can promote the expression of ROS-related proteins NOX2, NOX4, p-Src, and downstream proteins p-FAK, p-ROCK1/2, p-Cofilin-1, F-actin and inhibit the expression of p-SHP2 protein. Correction experiments showed that LIMK1 regulated the expression of p-FAK and p-Cofilin-1 proteins by regulating ROS and p-Src. Through the detection of cervical cancer cell functions, it was found that the activation of ROS and p-Src induced by LIMK1 is an early event that promotes the migration, proliferation, and invasion of cervical cancer cells.
CONCLUSIONS: LIMK1 promotes the expression of F-actin and promotes the development of cervical cancer by regulating the oxidative stress/Src-mediated p-FAK/p-ROCK1/2/p-Cofilin-1 pathway.
方法:LIMK1在调节生长中的生物学作用,入侵,在SiHa研究了宫颈癌的转移,CaSki细胞和裸鼠肿瘤模型。HE染色评价LIMK1在宫颈癌生长中的作用。LIMK1在入侵中的作用,转移,通过细胞划痕评估宫颈癌的增殖,Transwell,和单克隆实验。LIMK1、ROS、和Src通过Western印迹评价。通过细胞功能试验评价了调节ROS和p-Src表达对LIMK1在宫颈癌细胞迁移/侵袭和增殖中的作用。
结果:LIMK1过表达促进裸鼠肿瘤生长。细胞划痕,Transwell,单克隆实验表明LIMK1促进了入侵,转移,和宫颈癌细胞的增殖。Westernblotting提示LIMK1可促进ROS相关蛋白NOX2、NOX4、p-Src、和下游蛋白p-FAK,p-ROCK1/2、p-Cofilin-1、F-肌动蛋白和抑制p-SHP2蛋白的表达。校正实验表明,LIMK1通过调节ROS和p-Src调节p-FAK和p-Cofilin-1蛋白的表达。通过检测宫颈癌细胞的功能,发现LIMK1诱导的ROS和p-Src的激活是促进迁移的早期事件,扩散,和宫颈癌细胞的侵袭。
结论:LIMK1通过调节氧化应激/Src介导的p-FAK/p-ROCK1/2/p-Cofilin-1通路,促进F-actin的表达,促进宫颈癌的发生发展。