LRP1在肺癌细胞中的表达水平及其生物学功能研究  

The expression levels and biological function of LRP1 in lung cancer cells

作　　者：王海强 张天翼 张卫锋 尹迅亮 周勇安 赵正维 WANG Haiqiang;ZHANG Tianyi;ZHANG Weifeng;YIN Xunliang;ZHOU Yong'an;ZHAO Zhengwei(Department of Thoracic Surgery,the Second Affiliated Hospital of Air Force Medical University,Shaanxi Xi'an 710038,China.)

机构地区：[1]空军军医大学第二附属医院胸腔外科,陕西西安710038

出　　处：《现代肿瘤医学》2024年第19期3643-3647,共5页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81402411)。

摘　　要：目的:探讨低密度脂蛋白受体相关蛋白1(low density lipoprotein receptor-related protein 1,LRP1)在肺癌细胞中的表达水平及其生物学意义。方法:常规培养人类肺癌A549细胞,RT-PCR法检测A549细胞中LRP1的mRNA水平。构建针对LRP1的siRNA感染细胞,利用绿色荧光标记载体,通过荧光显微镜观察感染效率,利用RT-PCR检测敲减效率,Western blot检测LRP1表达水平变化。选用A549/LRP1 siRNA细胞系进行功能实验,利用CCK-8实验检测细胞增殖能力的变化,划痕实验检测细胞迁移能力的变化,Transwell侵袭小室实验检测细胞侵袭能力的变化。结果:从定量PCR结果可以看出,A549细胞中,相对于NC组,KD组LRP1基因敲减效率为77.7%,KD组细胞中LRP1的mRNA水平明显低于NC组,差异具有统计学意义(P<0.05);Western blot检测结果显示,KD组LRP1表达水平相对于NC组显著下调。CCK-8检测结果表明相对于NC组,KD组于Day 5的细胞增殖倍数显著下降,差异具有统计学意义(P<0.05)。划痕实验结果显示,相比NC组,KD组细胞划痕8小时、24小时迁移率明显降低,差异具有统计学意义(P<0.05)。Transwell侵袭实验结果显示,在侵袭小室内孵育24 h后KD组细胞侵袭转移率明显降低,差异具有统计学意义(P<0.05)。结论:LRP1对于肺癌A549细胞的生物学功能起着重要作用。下调LRP1能够抑制A549细胞的增殖、迁移及侵袭能力。对于LRP1分子上下游信号通路研究,在肺癌的诊断与治疗领域有一定的临床参考价值。Objective:To explore the expression levels and biological significance of low density lipoprotein receptor-related protein 1(LRP1)in lung cancer cells.Methods:Conventional culture of human lung cancer A549 cells,and RT-PCR method to detect the mRNA level of LRP1 in A549 cells.To construct siRNA infected cells targeting LRP1,using green fluorescent labeling vectors,observe infection efficiency through fluorescence microscopy,detect knockdown efficiency using RT-PCR,and detect changes in LRP1 expression levels using Western blot.A549/LRP1 siRNA cell lines were selected for functional experiments,and CCK-8 assay was used to detect changes in cell proliferation ability,scratch assay was used to detect changes in cell migration ability,and Transwell invasion chamber assay was used to detect changes in cell invasion ability.Results:From the quantitative PCR results,it can be seen that in A549 cells,compared to the NC group,the LRP1 gene knockout efficiency in the KD group was 77.7%.The mRNA level of LRP1 in the KD group cells was significantly lower than that in the NC group,and the difference was statistically significant(P<0.05).Western blot analysis showed that the expression level of LRP1 in the KD group was significantly downregulated compared to the NC group.The CCK-8 detection results showed that compared to the NC group,the KD group had a significant decrease in cell proliferation multiple on Day 5,and the difference was statistically significant(P<0.05).The scratch experiment results showed that compared with the NC group,the KD group showed a significant decrease in cell migration rates after 8 and 24 hours of scratch,with statistical significance(P<0.05).The results of Transwell invasion experiment showed that the invasion and metastasis rate of KD group cells significantly decreased after 24 hours of incubation in the invasion chamber,and the difference was statistically significant(P<0.05).Conclusion:LRP1 plays an important role in the biological function of lung cancer A549 cells.Downregulation of LRP1 ca

关 键 词：肺癌 A549细胞 LRP1 增殖 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]