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目的 探讨辛伐他汀通过上调Krüppel样因子4(KLF4)抑制丝裂原活化蛋白激酶（MAPK）/细胞外调节蛋白激酶（ERK）信号通路，进而抑制胰腺神经内分泌肿瘤（p NENs）细胞QGP-1增殖的作用机制。方法 通过RNA测序筛选辛伐他汀处理后的差异表达基因；利用肿瘤免疫评估资源（TIMER2.0）和基因表达谱交互分析平台2(GEPIA2)数据库分析KLF4在多种癌症中的表达模式及其与患者预后的关系。采用实时荧光定量PCR(RT-qPCR)和蛋白质免疫印迹法（Western blot）验证辛伐他汀对KLF4在转录与蛋白水平表达的影响；在KLF4表达较低的QGP-1细胞中构建KLF4过表达质粒，通过细胞计数试剂盒-8(CCK-8)、Ed U实验及平板克隆形成实验检测KLF4过表达对细胞增殖的影响；进一步通过Western blot检测MAPK/ERK通路关键蛋白水平的变化。结果 RNA-seq测序结果显示，经辛伐他汀处理后KLF4表达显著上调。生物信息学平台数据显示，KLF4在多种肿瘤组织中低表达，且其高表达与多种肿瘤的良好预后呈正相关（P<0.05）。RT-qPCR与Western blot实验结果显示，经辛伐他汀处理后，QGP-1细胞中KLF4在m RNA和蛋白水平均呈高表达（P<0.05）。Western blot结果证实KLF4在稳转株中成功过表达（P<0.001）。CCK-8、平板克隆形成实验和Ed U实验结果均显示KLF4过表达可显著抑制QGP-1细胞增殖。Western blot检测进一步证实KLF4过表达显著降低了磷酸化ERK(p ERK)水平（P<0.001）。结论辛伐他汀通过上调KLF4抑制MAPK/ERK信号通路活化，从而发挥抑制p NENs细胞增殖的作用。
Abstract：Objective To investigate the anti-tumor mechanism of simvastatin in pancreatic neuroendocrine neoplasms(pNENs) through regulating Krüppel-like factor 4(KLF4) and subsequently suppressing the mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase(ERK) signaling pathway. Methods Differentially expressed genes after simvastatin treatment were screened by RNA sequencing. The expression pattern of KLF4 in various cancers and its correlation with patient prognosis were analyzed using the TIMER2.0 and GEPIA2 databases. Reverse transcription-quantitative PCR(RT-qPCR) and Western blot were performed to validate the effect of simvastatin on KLF4 expression at transcriptional and protein levels. In QGP-1 cells with low endogenous KLF4 expression, a KLF4-overexpressing plasmid was constructed. Cell proliferation was assessed by CCK-8 assay, EdU incorporation assay, and colony formation assay. Western blot was further used to examine changes in key proteins of the MAPK/ERK pathway. Results RNA-seq results showed that KLF4 expression was upregulated after simvastatin treatment. Bioinformatics analyses indicated that KLF4 was lowly expressed in multiple tumor tissues, and its high expression was positively correlated with favorable prognosis in several cancers(P<0.05). Both RT-qPCR and Western blot confirmed that simvastatin treatment significantly increased KLF4 expression at mRNA and protein levels(P<0.05). Successful overexpression of KLF4 in stable transfection cells was verified by Western blot(P<0.001). CCK-8, colony formation, and EdU assays consistently demonstrated that KLF4 overexpression significantly suppressed the proliferation of QGP-1 cells. Furthermore, Western blot analysis revealed that KLF4 overexpression markedly reduced phosphorylated ERK(pERK) levels(P<0.001). Conclusion Simvastatin exerts anti-tumor effects in pNENs by upregulating KLF4 expression and inhibiting the MAPK/ERK signaling pathway.
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基本信息:
中图分类号:R735.9
引用信息:
[1]施晓婷,周璐,叶木杰,等.辛伐他汀通过调控KLF4介导的MAPK/ERK通路抑制胰腺神经内分泌肿瘤细胞增殖的研究[J].临床肿瘤学杂志,2026,31(02):134-140.
2026-02-28
2026-02-28