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三白草酮调控Hippo/YAP信号通路抑制非小细胞肺癌细胞的增殖和迁移

Saururus chinensis tannin inhibits proliferation and migration of non-small cell lung cancer cells by regulating Hippo/YAP signaling pathway

    摘要
  • 摘要:
    目的 探讨三白草酮通过调控Hippo/YAP信号通路对非小细胞肺癌(NSCLC)细胞增殖和迁移的影响。
    方法 使用不同浓度的三白草酮处理人NSCLC细胞系A549。采用CCK-8法检测细胞存活率，筛选药物浓度。将NSCLC细胞随机分为对照组和三白草酮低浓度组、三白草酮中浓度组、三白草酮高浓度组以及三白草酮高浓度+YAP激活剂(XMU-MP-1)组。使用CCK-8法、平板克隆实验、划痕实验、Transwell实验分别检测细胞存活率、细胞克隆形成情况、细胞迁移能力、细胞侵袭能力。采用蛋白质印迹检测迁移侵袭相关蛋白E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、snail及Hippo/YAP信号通路相关蛋白Yes相关蛋白(YAP)、磷酸化Yes相关蛋白(p-YAP)、结缔组织生长因子(CTGF)、半胱氨酸丰富血管生成诱导因子61(CYR61)的表达水平。
    结果 与对照组比较，三白草酮低浓度组、三白草酮中浓度组、三白草酮高浓度组的细胞存活率、克隆细胞数量、细胞迁移和侵袭能力、N-cadherin和snail蛋白表达水平以及YAP、CTGF、CYR61蛋白表达水平降低或减少, E-cadherin及p-YAP蛋白表达水平升高，差异有统计学意义(P＜0.05)，其中三白草酮高浓度组的效果最好。与三白草酮高浓度组比较，三白草酮高浓度+YAP激活剂组的细胞存活率、克隆细胞数量、细胞迁移和侵袭能力、N-cadherin和snail蛋白表达水平以及YAP、CTGF、CYR61蛋白表达水平升高或增加, E-cadherin及p-YAP蛋白表达水平降低，差异有统计学意义(P＜0.05)。
    结论 三白草酮可能通过调控Hippo/YAP信号通路，降低NSCLC细胞存活率，抑制其增殖、迁移和侵袭能力。

     

    Abstract:
    Objective To explore the effect of saururus chinensis tannin on proliferation and migration of non-small cell lung cancer (NSCLC) cells by regulating Hippo/YAP signaling pathway.
    Methods Human NSCLC cell line A549 was treated with different concentrations of saururus chinensis tannin. Cell viability was detected by CCK-8 assay to screen appropriate drug concentrations. NSCLC cells were randomly divided into control group, low-concentration saururus chinensis tannin group, medium-concentration saururus chinensis tannin group, high-concentration saururus chinensis tannin group and high-concentration saururus chinensis tannin+YAP activator (XMU-MP-1) group. Cell viability, colony formation capacity, migration ability and invasion ability were evaluated using the CCK-8 assay, plate colony formation assay, wound healing assay and transwell assay, respectively. Expression levels of proteins associated with migration and invasionE-cadherin, N-cadherin and snail and proteins related to the Hippo/YAP signaling pathway Yes-associated protein (YAP), phosphorylated Yes-associated protein (p-YAP), connective tissue growth factor (CTGF) and cysteine-rich angiogenic inducer 61 (CYR61)were detected by western blot analysis.
    Results Compared with the control group, the cell viability, the number of colony cells, migration and invasion abilities, as well as the expression levels of N-cadherin, snail, YAP, CTGF and CYR61 proteins in the low-concentration saururus chinensis tannin group, medium-concentration saururus chinensis tannin group and high-concentration saururus chinensis tannin group were significantly decreased or reduced, and the expression levels of E-cadherin and p-YAP proteins were significantly increased (P < 0.05), with the most pronounced effects observed in the high-concentration saururus chinensis tannin group. Compared with the high-concentration saururus chinensis tannin group, the cell viability, the number of colony cells, migration and invasion abilities, as well as the expression levels of N-cadherin, snail, YAP, CTGF and CYR61 in the high-concentration saururus chinensis tannin+YAP activator group were significantly increased or enhanced, while the expression levels of E-cadherin and p-YAP were significantly decreased (P < 0.05).
    Conclusion Saururus chinensis tannin may reduce the survival rate of NSCLC cells and inhibit their proliferation, migration and invasion abilities by regulating the Hippo/YAP signaling pathway.

     

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