干扰LASP1基因表达对胃癌细胞BGC823增殖、迁移及侵袭能力的影响

发布时间：2018-09-08 12:15

【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(P0.05);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(P0.05);在干扰组细胞内的F-actin聚合量比对照组减少(P0.05);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。
[Abstract]:Background & AIM: LASP1 is a novel actin binding protein involved in the regulation of cytoskeleton recombination and cell migration. It is highly expressed in various malignant tumors and is closely related to tumor proliferation invasion and metastasis. However, the role of LASP1 in the occurrence and development of gastric cancer is rarely reported. The aim of this study was to investigate the role and molecular mechanism of LASP1 in the proliferation and invasion of gastric cancer. Methods: real-time fluorescence quantitative polymerase chain reaction (real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR) and Western blot (Western blot) were used to detect the expression of LASP1 in different gastric cancer cell lines. RNA interference technique was used to inhibit the expression of LASP1 in BGC823 cells. RTFQ-PCR and Western blot were used to detect the expression of LASP1 after transfection. The proliferation, migration and invasion of transfected cells were detected by in vitro proliferation, migration and invasion experiments. The aggregation ability of F-actin was detected by F-actin polymerization assay. Western blot was used to detect the phosphorylation of Akt in BGC823 cells before and after transfection. Results the expression of m RNA and protein in the interfering group was significantly lower than that in the control group after the LASP1 gene was silenced by the high expression of .RNAi technique in BGC823 cells of gastric cancer. Cell proliferation test showed that the cell proliferation rate in the interference group was lower than that in the control group (P0.05). The ability of migration and invasion were significantly decreased in the interference group (P0.05); the amount of F-actin aggregation in the interference group was lower than that in the control group (P0.05); the phosphorylation of Akt was inhibited in the interference group. Conclusion the decreased expression of WASP1 can inhibit the proliferation of gastric cancer BGC823 cells in vitro. The migration and invasion ability of LASP1 may affect the invasion and metastasis of gastric cancer cells by regulating the aggregation of F-actin and the phosphorylation of Akt.
【作者单位】：潍坊医学院临床病理系神经疾病与再生修复实验室;
【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)
【分类号】：R735.2

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