Cheng Xudong, Gu Junfei, Zhang Minghua, Yuan Jiarui, Zhao Bingjie, Jiang Jun, Jia Xiaobin
College of Pharmacy, Nanjing University of Chinese Medicine, Jiangsu 210046, China; Key Laboratory of New Drug Delivery System of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu 210028, China.
Key Laboratory of New Drug Delivery System of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu 210028, China; College of Pharmacy, Jiangsu University, Jiangsu 212013, China.
Int Immunopharmacol. 2014 Nov;23(1):304-13. doi: 10.1016/j.intimp.2014.08.027. Epub 2014 Sep 16.
The migration and invasion characteristics that are related to inflammatory response play important roles in the development of lung cancer. Astagaloside IV (AS-IV), an effective saponin component isolated from Astragali Radix, has been reported to inhibit metastasis of tumor cells. However, little is known about the underlying mechanism of AS-IV on inhibiting the migration and invasion characteristics of lung cancer cells. In the present study, cell proliferation was assessed by MTT colorimetric assay. Wound-healing assay and transwell chambers assay were used to detect the effects of AS-IV on the migration capacity and invasiveness of A549 cells. Metastasis-related bio-markers expressions were detected by Western blot analysis. Levels of inflammatory factors including transforming growth factor-β1 (TGF-β1), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in cell supernatant were tested by enzyme linked immunosorbent assay (ELISA). The expressions of PKC-α, ERK1/2 and NF-κB were analyzed by Western blot analysis. The results showed that the migration and invasion ability of A549 has been suppressed in presence of AS-IV. The levels of MMP-2, MMP-9 and integrin β1 were decreased significantly, whereas E-cadherin was increased by the treatment of different concentrations AS-IV. Furthermore, AS-IV also significantly decreased TGF-β1, TNF-α and IL-6 levels. Interestingly, PKC pathway inhibitor AEB071 (Sotrastaurin) (0.1 μM) or ERK inhibitor U0126 (1 μM) or NF-κB inhibitor PDTC (1 μM) could affect suppression of AS-IV on cell invasion, at least partially. Our results suggested that the migration and invasion of AS-IV in A549 cells might be related to the PKC-α-ERK1/2-NF-κB pathway. The result indicated that AS-IV could be used as a candidate for the inhibition of metastasis of human lung cancer.
与炎症反应相关的迁移和侵袭特性在肺癌发展中起重要作用。黄芪甲苷IV（AS-IV）是从黄芪中分离出的一种有效皂苷成分，据报道可抑制肿瘤细胞转移。然而，关于AS-IV抑制肺癌细胞迁移和侵袭特性的潜在机制知之甚少。在本研究中，通过MTT比色法评估细胞增殖。采用伤口愈合试验和Transwell小室试验检测AS-IV对A549细胞迁移能力和侵袭性的影响。通过蛋白质免疫印迹分析检测转移相关生物标志物的表达。采用酶联免疫吸附测定（ELISA）检测细胞上清液中包括转化生长因子-β1（TGF-β1）、肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）在内的炎症因子水平。通过蛋白质免疫印迹分析PKC-α、ERK1/2和NF-κB的表达。结果表明，AS-IV存在时A549的迁移和侵袭能力受到抑制。不同浓度AS-IV处理后，MMP-2、MMP-9和整合素β1水平显著降低，而E-钙黏蛋白增加。此外，AS-IV还显著降低TGF-β1、TNF-α和IL-6水平。有趣的是，PKC通路抑制剂AEB071（索拉苏林）（0.1 μM）或ERK抑制剂U0126（1 μM）或NF-κB抑制剂PDTC（1 μM）至少部分影响AS-IV对细胞侵袭的抑制作用。我们的结果表明，AS-IV在A549细胞中的迁移和侵袭可能与PKC-α-ERK1/2-NF-κB通路有关。结果表明，AS-IV可作为抑制人肺癌转移的候选药物。
