Effect of Feiyanning Formula on Invasion and Metastasis of Lung Cancer  Inhibiting TGF- Mediated Bone Metastasis Related PTHrP/RANKL Signaling Pathway

ZHANG Cheng-cheng; JI Qing; YANG Yue; WU Xin-nan; QIN Jing-ru; XIE Yu; LI Qi; WANG Zhong-qi

doi:10.13422/j.cnki.syfjx.2018030132

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Effect of Feiyanning Formula on Invasion and Metastasis of Lung Cancer Inhibiting TGF- Mediated Bone Metastasis Related PTHrP/RANKL Signaling Pathway

更新时间：2024-01-04

- Effect of Feiyanning Formula on Invasion and Metastasis of Lung Cancer Inhibiting TGF- Mediated Bone Metastasis Related PTHrP/RANKL Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 3, Pages: 132-136(2018)
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- DOI：10.13422/j.cnki.syfjx.2018030132
  CLC： R285
- Published：2018
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ZHANG Cheng-cheng, JI Qing, YANG Yue, et al. Effect of Feiyanning Formula on Invasion and Metastasis of Lung Cancer Inhibiting TGF- Mediated Bone Metastasis Related PTHrP/RANKL Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(3): 132-136.
DOI：

ZHANG Cheng-cheng, JI Qing, YANG Yue, et al. Effect of Feiyanning Formula on Invasion and Metastasis of Lung Cancer Inhibiting TGF- Mediated Bone Metastasis Related PTHrP/RANKL Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(3): 132-136. DOI： 10.13422/j.cnki.syfjx.2018030132.

摘要

目的：探讨肺岩宁方通过调控转化生长因子-β（transforming growth factor-β，TGF-β）介导的骨转移相关甲状旁腺激素相关蛋白（parathyroid hormone-related protein，PTHrP）/NF-κB受体活化因子配体（receptor or activator of NF-κB ligand，RANKL）信号通路抗肺癌细胞侵袭转移的作用机制。方法：体外培养A549肺癌细胞，用不同终浓度肺岩宁方进行处理，未加肺岩宁方为空白组。分别采用cell counting kit-8（CCK-8）检测肺岩宁方对A549细胞增殖抑制作用，计算半数浓度抑制率（IC50）；采用转移小室（transwell）侵袭实验、划痕实验检测肺岩宁方对A549细胞体外侵袭及迁移的抑制作用；以及蛋白免疫印迹法（Western blot）检测TGF-β，PTHrP，RANKL蛋白表达。结果：肺岩宁方作用于A549肺癌细胞48 h后，与空白组相比，肺岩宁方（12.5，25，50 mg·L-1）组穿膜细胞明显减少（P<0.05，P<0.01）；肺岩宁方（12.5，25，50 mg·L-1）组的划痕距离较空白组明显降低（P<0.05，P<0.01）；且肺岩宁方（25，50 mg·L-1）组作用于A549细胞后，TGF-β蛋白表达明显下调（P<0.05），其对应下游靶基因PTHrP及RANKL蛋白也发生明显表达下调（P<0.05）。结论：肺岩宁方可抑制A549肺癌细胞的侵袭迁移能力，直接下调TGF-β基因及其下游靶基因PTHrP，RANKL蛋白表达，且具有浓度相关性。本研究证实了肺岩宁方通过调控TGF-β介导的骨转移相关PTHrP/RANKL信号通路，以达到抗肺癌侵袭转移的作用机制，这为肺癌临床靶向治疗提供了一定的实验依据，也显示了肺岩宁方作为抗肺癌侵袭转移药物广泛的应用前景。

Abstract

Objective:To investigate the effect mechanism of Feiyanning formula on invasion and metastasis of lung cancer cells via inhibiting transforming growth factor-β (TGF-β) mediated bone metastasis related parathyroid hormone-related protein (PTHrP)/receptor or activator of NF-κB ligand (RANKL) signaling pathway. Method:A549 cells were cultured in vitro and treated with Feiyanning formula with various concentrations

with no Feiyanning formula as blank group. The anti-proliferation effects and the 50% inhibitory concentration (IC50) of Feiyanning formula was measured by cell counting kit-8 (CCK-8) method. The inhibiting effect against invasion and migration ability of A549 cells was detected by transwell assay and scratch test. Western blot was used to detect the protein expression of TGF-β

parathyroid hormone-related protein (PTHrP) and receptor or activator of NF-κB ligand (RANKL). Result:After Feiyanning formula effected on A549 lung cancer cells for 48 h

as compared with blank control group

the transmembrane cells in the low

medium and high dose Feiyanning formula (12.5

50 mg·L-1) groups were obviously reduced (P<0.05

P<0.01)

and the scratch distance in Feiyanning formula groups was also significantly shortened (P<0.05

P<0.01). Furthermore

medium and high dose Feiyanning formula group significantly down-regulated the expression of TGF-β (P<0.05) and its corresponding downstream target genes of PTHrP and RANKL (P<0.05). Conclusion:Feiyanning formula can directly down-regulate the protein expression of TGF-β and its downstream target genes PTHrP and RANKL

and inhibit the invasion and migration ability of A549 lung cancer cells in a concentration-dependent manner. This study confirmed the mechanism of Feiyanning formula on inhibiting the invasion and migration of lung cancer by regulating TGF-β mediated bone metastasis related PTHrP/RANKL signaling pathway

providing a certain experimental basis for the clinical target therapy of lung cancer

suggesting that Feiyanning formula could be a potential candidate in the treatment of lung cancer cell bone metastasis.

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