摘要
背景与目的:LASP1是一种新的肌动蛋白结合蛋白,参与细胞骨架的重组调控及细胞迁移,在多种恶性肿瘤中高表达,并与肿瘤的增殖、侵袭和转移密切相关。但目前LASP1在胃癌发生、发展中的作用少见报道。该研究旨在探讨LASP1在胃癌增殖和侵袭中的作用及分子机制。方法:利用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)和蛋白[质]印迹法(Western blot)检测LASP1在不同胃癌细胞系中的表达,应用RNA干扰技术抑制BGC823细胞中LASP1的表达,采用RTFQ-PCR和Western blot检测转染后LASP1的表达,应用体外增殖、迁移和侵袭实验检测转染后细胞的增殖、迁移和侵袭能力,通过F-actin聚合实验检测细胞的F-actin的聚合能力,采用Western blot检测转染前后BGC823细胞中Akt的磷酸化情况。结果:LASP1在胃癌BGC823细胞中高表达。RNA干扰技术沉默LASP1基因后,干扰组与对照组相比,LASP1的m RNA和蛋白表达水平均明显下降。细胞增殖实验显示,干扰组细胞的增殖率低于对照组(P<0.05);Transwell迁移和侵袭实验发现,与对照组相比,干扰组细胞的迁移和侵袭能力均明显降低(P<0.05);在干扰组细胞内的F-actin聚合量比对照组减少(P<0.05);在干扰组细胞内Akt磷酸化受到抑制。结论:LASP1的表达降低可以抑制胃癌BGC823细胞的体外增殖、迁移和侵袭能力,LASP1通过调节F-actin的聚合和Akt磷酸化从而影响胃癌细胞的侵袭、转移。
Background and purpose： LASP1 is a novel actin-binding protein and involved in the process of cytoskeleton reorganization and cell migration. Expression of LASP1 is significantly up-regulated in several malignant tumors and is closely related to the tumor proliferation, invasion and metastasis. However, the specific mechanism of LASP1 is not clear in gastric carcinogenesis. This study aimed to detect the effect and mechanism of LASP1 in the proliferation and invasion of gastric cancer cells. Methods： Real-time fluorescence quantitative polymerase chain reaction （RTFQ-PCR） and Western blot were employed to determine the expression of LASP1 in gastric cancer cell lines, as well as human normal gastric epithelial cell line GES-1. RNA interference （RNAi） was used to silence the expression of LASP1 in gastric cancer cell BGC823. RTFQ-PCR and Western blot were used to analyze the expression of LASP1 after siRNA transfection. In vitro proliferation, migration and invasion assay were performed to detect the effect of LASP1 on BGC823 cells. F-actin polymerization assay was used to detect F-actin recognition ability in cells. Western blot was used to analyze phosphorylation of Akt. Results： Overexpression of LASP1 was detected in human gastric cancer BGC823 cells. Down-regulated expression of LASP1 at both mRNA and protein level was detected after siRNA transfection. Cell proliferation assay showed that the growth rate was significantly reduced in RNAi group （P〈0.05）. In transwell assay, the cell migration and invasion were significantly decreased （P〈0.05）. F-actin content was reduced compared with the control group （P〈0.05）. The phosphorylation of Akt was significantly reduced after siRNA transfection. Conclusion： LASP1 was highly expressed in gastric cancer cells. Down-regulation of LASP1 suppressed the capacities of proliferation, migration and invasion of human gastric cancer BGC823 cells in vitro. LASP1 can induce migration and invasion of gastric cancer through regulation of F-actin polymerization and Akt phosphorylation.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2017年第11期847-852,共6页
China Oncology
基金
国家自然科学基金(81472365)
山东省自然科学基金(ZR2015HL091
ZR2014HM086
ZR2016HL20)
山东省医药卫生科技发展计划(2013WS0279
2014WS0182)
山东省高等学校科技计划(J15LK11)
