中药QHF复方抑制乳腺癌MCF-7细胞增殖、迁移、侵袭的机制研究

摘要：目的 研究中药QHF复方对乳腺癌MCF-7细胞增殖、迁移、侵袭的影响及其可能的作用机制,为QHF的临床应用提供理论依据。方法 体外培养人乳腺癌MCF-7细胞,以对数生长期的细胞为研究对象,予以不同浓度QHF复方及其他药物,CCK8法检测QHF复方对乳腺癌MCF-7细胞生长增殖的影响。细胞划痕实验和Transwell实验检测QHF复方对乳腺癌MCF-7细胞迁移、侵袭的影响。Western blot检测QHF复方对乳腺癌MCF-7细胞HGF/c-Met及其下游PI3K/Akt、MAPK/ERK信号通路的影响。结果 CCK8实验结果显示：中药QHF复方可不同程度抑制乳腺癌MCF-7细胞增殖且这种抑制作用可显著拮抗HGF对乳腺癌MCF-7细胞增殖的促进作用。细胞划痕实验和Transwell实验结果显示：中药QHF复方能明显抑制MCF-7细胞迁移、侵袭且这种抑制作用可拮抗HGF 对MCF-7细胞迁移、侵袭的促进作用。Western blot实验结果显示：中药QHF复方可明显抑制乳腺癌MCF-7细胞HGF、p-c-Met、p-Akt、p-ERK、VEGF、MMP-2、MMP-9的表达。结论 中药QHF复方抑制乳腺癌MCF-7细胞的增殖、迁移和侵袭的机制可能与抑制乳腺癌MCF-7细胞异常活化的HGF/c-Met及其下游PI3K/Akt、MAPK/ERK信号通路有关。

Mechanism of Chinese herbal formula QHF against breast cancer MCF-7 cells invasion and migration

Objective

To investigate the effects of Chinese herbal formula Qinghuofu (QHF) on the migration and invasion of breast cancer MCF-7 cells and its possible molecular mechanisms, thereby providing a theoretical basis to find effective anti-cancer medicine and therapeutic targets for the treatment of anti-migration and anti-invasion of breast cancer.

Methods

Breast cancer MCF-7 cells were treated with different QHF and other different reagents, CCK8 assay was used to detect the influence of the reagents on the proliferation of MCF-7 cells; Scrape migration and Transwell assay were used to quantitatively determine the migration and invasion effects of QHF and hepatocyte growth factor (HGF) on the MCF-7 cells. Subsequently, the c-Met inhibitor and its downstream ERK and PI3K inhibitors were used to investigate the relationship between the migration and invasion of MCF-7 cells, as well as its downstream MAPK/ERK and PI3K/Akt signaling pathways. The expression levels of HGF, c-Met, ERK, p-Akt, p-c-Met, p-ERK, p-Akt, MMP2, MMP9, and VEGF in breast cancer MCF-7 cells treated with QHF and other reagents were also examined.

Results

The result indicated that formula QHF not only significantly inhibited the proliferation of MCF-7 cells, but also significantly suppressed the effects of HGF (40?ng/mL) on the proliferation and movement of MCF-7 cells, reducing the ability of the cells to invade and migrate. Western blot analysis indicated that QHF and c-Met inhibitor significantly decreased the expression of p-c-Met, p-ERK1, p-ERK2, p-Akt, MMP-2, MMP-9, and VEGF, while HGF significantly increased the expression of p-c-Met in MCF-7 cells; c-Met downstream ERK and PI3K inhibitors also significantly decreased the expression of MMP-2, MMP-9, and VEGF in MCF-7 cells; But the difference among c-Met, PI3K, ERK, and QHF group were not statistically significant.

Conclusion

QHF can prevent the proliferation, migration, and invasion of MCF-7 cells by inhibiting the HGF/c-Met and its downstream PI3K/Akt and MAPK/ERK signaling pathways; Thereby down-regulating the expression of HGF, p-Met, p-ERK1, p-ERK2, p-Akt, MMP-2, MMP-9, and VEGF.