TPM4 promotes cell migration by modulating F-actin formation in lung cancer
TPM4 promotes cell migration by modulating F-actin formation in lung cancer
复制标题
TPM4 通过调节肺癌中 F-肌动蛋白的形成促进细胞迁移
DOI：
10.2147/ott.s198542
复制
发表时间：
2019-01-01
影响因子：
4
通讯作者：
Wang, Ziyu
中科院分区：
文献类型：
--
作者：
Zhao, Xiaoting;Jiang, Mei;Wang, Ziyu
背景：原肌球蛋白4(TPM4)是肌动蛋白结合蛋白原肌球蛋白家族的成员。TPM4可作为卵巢癌、乳腺癌、结肠癌、角化棘皮瘤和食管鳞癌的潜在检测指标。本文对TPM4在肺癌细胞系中的功能进行了研究。材料和方法：采用CRISPR/Cas9技术构建TPM4基因敲除细胞。在TPM4基因敲除细胞的基础上构建了TPM4过表达细胞。MTS法检测细胞生长能力。用Transwell实验和创伤划痕实验检测细胞的运动能力。用免疫印迹法检测肺癌细胞的蛋白质水平。采用免疫荧光技术对F-肌动蛋白的结构进行图像分析。结果：成功建立了TPM4下调和TPM4上调的细胞模型。在TPM4上调的细胞中，TPM4抑制细胞的运动，而促进细胞的迁移。但TPM4不参与细胞增殖和EMT进展。TPM4的表达受到抑制，导致微丝解聚。当TPM4表达上调时，F-肌动蛋白组装增加。结论：TPM4通过直接改变肌动蛋白细胞骨架而促进细胞运动。
Background: Tropomyosin 4 (TPM4) is a member of the tropomyosin family of actin-binding proteins. Abnormal level of TPM4 is found in several cancers, and TPM4 is considered as a potential detecting marker for ovarian cancer, breast cancer, colon cancer, keratoacanthoma and esophageal squamous cell carcinoma. In this paper, the function of TPM4 in lung cancer cell lines was determined. Materials and methods: TPM4 knockout cells were constructed by CRISPR/CAS9 technique. TPM4 overexpression cells were also constructed based on TPM4 knockout cells. Cell growth ability was detected by MTS assay. The potency of cell motility was investigated using transwell assay and wound scratch assay. The protein levels in lung cancer cells were determined by western-blot. Immunofluorescence technique was used to image the structure of F-actin. Results: As a result, TPM4 downregulation and TPM4 upregulation cell models were obtained successfully. Cell motility was inhibited by the suppression of TPM4 while cell migration was enhanced in TPM4 upregulated cells. But TPM4 was not involved in cell proliferation and EMT progression. Microfilaments were depolymerized result from the suppression of TPM4 expression. And F-actin assembly was increased when TPM4 was upregulated. Conclusion: In summary, TPM4 was able to promote cell motility by altering the actin cytoskeleton directly.