骨架蛋白MYH9和Palladin在肺癌发生发展中的联合作用和临床价值

The combined effect and clinical value of cytoskeletal proteins MYH9 and Palladin in lung cancer

目的:研究骨架蛋白MYH9和Palladin对肺癌患者预后和恶性表型的联合作用及临床价值。方法:从癌症基因组图谱(TCGA)数据库下载肺癌转录组数据以及相关临床信息。利用生物信息学方法,分析MYH9和Palladin对肺癌患者预后及其与临床病理特征的关系,进一步分析两者对肺癌预后的联合作用。同时构建双分子敲降和单分子敲降的NCI-H460细胞,分别采用CCK-8法检测双分子敲降对肺癌细胞增殖和耐药的影响,Transwell实验检测双分子联合敲降对肺癌细胞迁移和侵袭的影响,甲基纤维素成球实验检测双分子敲降对肺癌细胞自我更新的作用。最后通过组织芯片进一步验证MYH9和Palladin联合在肺癌中的表达及其对患者预后的影响。结果:生物信息学分析结果显示MYH9和Palladin在肺癌组织中均呈现高表达,相比于低表达组患者,MYH9和Palladin高表达组患者的预后以及临床分期更差,其中双分子均高表达组肺癌患者的预后最差(P<0.01)。细胞生物学实验结果表明双分子敲降组肺癌细胞增殖、耐药性、迁移、侵袭及自我更新能力均明显低于单分子敲降组和对照组细胞(P<0.01)。组织芯片结果进一步证实了相比于癌旁组织,MYH9和Palladin在肺癌组织中均呈现高表达,与低表达组相比,MYH9和Palladin高表达组的肺癌患者预后更差(P<0.01),且双分子均高表达组肺癌患者的临床预后最差。结论:MYH9和Palladin的联合作用对肺癌细胞干性相关特征的影响显著高于其单独作用,MYH9和Palladin双分子联合对肺癌的临床预后作用更显著,这表明MYH9和Palladin有可能成为靶向非小细胞肺癌的联合靶标分子。

OBJECTIVE:This study aimed to investigate the combined effects and clinical value of the cytoskeletal proteins MYH9 and Palladin on prognosis and malignant phenotypes in lung cancer.METHODS:Transcriptomic data of lung cancer,along with relevant clinical information,were downloaded from The Cancer Genome Atlas(TCGA)database.Bioinformatics analyses were conducted to assess the influence of MYH9 and Palladin on lung cancer prognosis and clinical staging,and to further analyze their combined effects on lung cancer prognosis.NCI-H460 cells with double and single knockdowns of MYH9 and Palladin were constructed.The effects of these gene knockdowns on lung cancer cell proliferation,drug resistance,migration,invasion,and self-renewal were evaluated using the CCK-8 assay,transwell assays,and methylcellulose sphere formation assay,respectively.Finally,the expression of MYH9 and Palladin in lung cancer and their influence on patient prognosis were further verified by tissue microarray.RESULTS:The analysis of biological information revealed that MYH9 and Palladin were markedly expressed in lung cancer tissues.Patients in the high expression group of MYH9 and Palladin exhibited a worse prognosis and clinical stage compared to those in the low expression group.Furthermore,the combined high expression group demonstrated the poorest prognosis for lung cancer patients(P<0.01).Results from the cell cuture experiment demonstrated that the proliferation,drug resistance,migration,invasion,and self-renewal ability of lung cancer cells in the bimolecular knockout group were significantly lower than those in the single-molecule knockout group and the control cells(P<0.01).Results from the tissue microarray further confirmed that MYH9 and Palladin were highly expressed in lung cancer tissues in comparison to adjacent paracancerous tissues.In comparison to the low expression group,the prognosis of lung cancer patients in the high expression group of MYH9 and Palladin was worse,and the bimolecular combination of high expression group had a more significant effect on the clinical prognosis of lung cancer patients.CONCLUSION:The combined effect of MYH9 and Palladin on clinical prognosis in lung cancer was more pronounced than their individual effects.Additionally,their combined impact on stemness-related characteristics in lung cancer cells was significantly greater than that of either MYH9 or Palladin alone.These findings suggest that MYH9 and Palladin could potentially serve as combined therapeutic targets for non-small cell lung cancer.