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Received:March 19, 2021 Published Online:January 20, 2022
Received:March 19, 2021 Published Online:January 20, 2022
中文摘要: 目的 通过体外实验探讨非小细胞肺癌(NSCLC)A549细胞系的Vinculin表达对其生物学特征的影响。方法 购买人NSCLC A549细胞系,体外培养,分为对照组、空白载体组以及高表达组三组,高表达组A549细胞系转染Vinculin过表达载体,空白载体组A549细胞系转染空白载体。体外培养48 h后,应用RT-PCR检测各组A549细胞系Vinculin mRNA表达情况,应用MTT法检测细胞活性,Ki-67免疫荧光检测细胞增殖能力,Transwell培养体系Hoechst染色观察A549细胞系迁移侵袭能力。结果 RT-PCR检测结果显示,与对照组、空白载体组比较,高表达组Vinculin mRNA表达量明显增高(P<0.01);MTT法检测结果显示,与对照组、空白载体组比较,高表达组的OD值明显降低(P<0.01);Ki-67免疫荧光检测结果显示,与对照组、空白载体组比较,高表达组Ki-67+细胞数量明显减少(P<0.01);Transwell培养体系Hoeschst染色结果显示,与对照组、空白载体组比较,高表达组迁移和侵袭的细胞数量明显减少(P<0.01)。上述各项指标组间比较差异均有统计学意义。结论 高表达Vinculin能够抑制NSCLC A549细胞系增殖、迁移以及侵袭能力,Vinculin可能是NSCLC诊治的潜在生物学靶点。
Abstract:Objective To explore the influences of Vinculin expression on the biological characteristics of non-small cell lung cancer(NSCLC) A549 cell lines through in-vitro experiments.
Methods After being purchased and cultured in vitro,human NSCLC A549 cell lines were divided into control group, blank vector group(transfected with blank vector) and high expression group(transfected with Vinculin overexpression vector). After in-vitro culture for 48 hours,RT-PCR was used to detect the expression of Vinculin mRNA in A549 cells, MTT was used to measure the cell viability, Ki-67 immunofluorescence was used to determine the cell proliferation, and Transwell culture system and Hoechst staining was used to observe the migration and invasion ability of A549 cells.
Results RT-PCR showed that the expression of Vinculin mRNA in high expression group significantly increased compared with control group and blank vector group (P<0.01). MTT assay showed that the OD value in high expression group was significantly lower than that in control group and blank vector group (P<0.01). Ki-67 immunofluorescence test showed that the number of Ki-67+ cells in high expression group significantly decreased compared with control group and blank vector group (P<0.01). Transwell culture system and Hoeschst staining results showed that the number of migrated and invaded cells in high expression group was significantly lower than those in control group and blank vector group (P<0.01).
Conclusions Overexpression of Vinculin can inhibit the proliferation, migration and invasion of NSCLC A549 cell line. Vinculin may be a potential biological target for the diagnosis and treatment of NSCLC.
文章编号: 中图分类号:R734.2 文献标志码:A
基金项目:南通市科技计划项目(MSZ1844)
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