Wang Zi-Ling, Guo Yu-Jie, Zhu Yun-Yun, Chen Le, Wu Ting, Liu Da-Hui, Huang Bi-Sheng, DU Hong-Zhi
Traditional Chinese Medicine Resource Center, Hubei University of Chinese Medicine Wuhan 430065, China.
Traditional Chinese Medicine Resource Center, Hubei University of Chinese Medicine Wuhan 430065, China Hubei Provincial Key Laboratory of Traditional Chinese Medicine Resource and Traditional Chinese Medicine Chemistry, Hubei University of Chinese Medicine Wuhan 430065, China.
Zhongguo Zhong Yao Za Zhi. 2021 Oct;46(20):5362-5371. doi: 10.19540/j.cnki.cjcmm.20210628.701.
The present study explored the effects and its underlying mechanisms of four active fractions of Camellia nitidissima(leaf polyphenols, leaf saponins, flower polyphenols, and flower saponins in C. nitidissima) in inhibiting the proliferation and migration of non-small cell lung cancer(NSCLC) by suppressing the epidermal growth factor receptor(EGFR). MTT assay was used to detect the effect of four active fractions on the proliferation of NCI-H1975 and HCC827 cells. Wound healing assay and Transwell assay were adopted to evaluate the effect of four active fractions on the migration of NSCLC. The effect of four active fractions on the enzyme activity of EGFR was detected. Molecular docking was carried out to explore the direct action capacity and action sites between representative components of the four active fractions and EGPR. Western blot assay was employed to investigate the effect of four active fractions on the protein expression in EGFR downstream signaling pathways. The results of the MTT assay indicated that the cell viability of NCI-H1975 and HCC827 cells was significantly inhibited by four active fractions at 50, 100, 150, and 200 μg·mL~(-1) in a dose-dependent manner. Wound healing assay and Transwell assay revealed that the migration of NCI-H1975 and HCC827 cells was significantly suppressed by four active fractions. In addition, the results of the protein activity assay showed that the enzyme activity of EGFR was significantly inhibited by four active fractions. The molecular docking results confirmed that various components in four active fractions possessed strong binding activity to EGFR enzymes. Western blot assay revealed that four active fractions down-regulated the protein expression of EGFR and its downstream signaling pathways. It is concluded that the four active fractions of C. nitidissima can inhibit NSCLC. The mechanism may be related to EGFR and its downstream signaling pathways. This study provides a new scientific basis for the clinical treatment of NSCLC with active fractions of C. nitidissima, which is of reference significance for further research on the anti-tumor mechanism of C. nitidissima.
本研究探讨了金花茶的四种活性成分(金花茶叶多酚、叶皂苷、花多酚和花皂苷)通过抑制表皮生长因子受体(EGFR)来抑制非小细胞肺癌(NSCLC)增殖和迁移的作用及其潜在机制。采用MTT法检测四种活性成分对NCI-H1975和HCC827细胞增殖的影响。采用划痕实验和Transwell实验评估四种活性成分对NSCLC迁移的影响。检测四种活性成分对EGFR酶活性的影响。进行分子对接以探究四种活性成分的代表性成分与EGPR之间的直接作用能力和作用位点。采用蛋白质免疫印迹法研究四种活性成分对EGFR下游信号通路中蛋白质表达的影响。MTT实验结果表明,四种活性成分在50、100、150和200μg·mL⁻¹浓度下均能显著抑制NCI-H1975和HCC827细胞的活力,且呈剂量依赖性。划痕实验和Transwell实验显示,四种活性成分显著抑制了NCI-H1975和HCC827细胞的迁移。此外,蛋白质活性检测结果表明,四种活性成分显著抑制了EGFR的酶活性。分子对接结果证实,四种活性成分中的各种成分对EGFR酶具有较强的结合活性。蛋白质免疫印迹法显示,四种活性成分下调了EGFR及其下游信号通路的蛋白质表达。结论是金花茶的四种活性成分可抑制NSCLC。其机制可能与EGFR及其下游信号通路有关。本研究为金花茶活性成分临床治疗NSCLC提供了新的科学依据,对进一步研究金花茶的抗肿瘤机制具有参考意义。