Shi He, Wang Yange, Yao Mengli, Zhang Dian, Fang Wenli, Zhou Ting, Gan Delu, Yue Shujun, Qian Husun, Chen Tingmei
Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
Transl Cancer Res. 2020 Dec;9(12):7596-7604. doi: 10.21037/tcr-20-3110.
Breast cancer is one of the most malignant tumors in the reproductive system and has a poor prognosis. Finding drugs with high efficiency, low side-effects, and low cost has become a research hotspot.
In the present study, we treated SK-BR-3 cells with different doses of honokiol. Crystal violet staining method was used to detect changes in the total number of living cells; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to detect the effect of honokiol on SK-BR-3 cell proliferation. Cell migration ability change was determined by wound healing assay. Cell invasion ability change was determined by Transwell migration assay. Flow cytometry was used to detect the apoptotic rate of SK-BR-3 cells, and Western blot was used to detect the expression levels of proliferation-associated protein (PCNA); migration- and invasion-related protein matrix metalloproteinase-2 (MMP-2); vimentin; apoptosis-related proteins Bcl-xl, caspase 3, and cleaved caspase 3 (CC3); and β-catenin and its downstream target molecule c-Myc.
Compared with the control group, different doses of honokiol have different degrees of inhibitory effects on cells, including proliferation and invasion and migration (P<0.01). After treatment with 50 or 60 µmol·L honokiol, the apoptotic rate of SK-BR-3 cells increased (both P<0.01); PCNA expression was significantly downregulated (P<0.01). Intracellular accumulation of apoptosis-related proteins Bcl-xl and caspase-3 decreased but C-C3 increased. We also found downregulation of MMP-2 expression, a protein related to invasion and migration (P<0.01), and a decrease in the expression levels of the Wnt/β-catenin signaling pathway-related proteins β-catenin and c-Myc (P<0.01).
Honokiol can promote the apoptosis of SK-BR-3 cells and can inhibit the proliferation, migration, and invasion of human breast cancer SK-BR-3 cells. The underlying mechanism may be through inhibiting the activation of the Wnt signaling pathway.
乳腺癌是生殖系统中最恶性的肿瘤之一，预后较差。寻找高效、低副作用且低成本的药物已成为研究热点。
在本研究中，我们用不同剂量的厚朴酚处理SK-BR-3细胞。采用结晶紫染色法检测活细胞总数的变化；采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法检测厚朴酚对SK-BR-3细胞增殖的影响。通过伤口愈合试验测定细胞迁移能力的变化。通过Transwell迁移试验测定细胞侵袭能力的变化。采用流式细胞术检测SK-BR-3细胞的凋亡率，采用蛋白质免疫印迹法检测增殖相关蛋白（PCNA）；迁移和侵袭相关蛋白基质金属蛋白酶-2（MMP-2）；波形蛋白；凋亡相关蛋白Bcl-xl、半胱天冬酶3和裂解的半胱天冬酶3（CC3）；以及β-连环蛋白及其下游靶分子c-Myc的表达水平。
与对照组相比，不同剂量的厚朴酚对细胞具有不同程度的抑制作用，包括增殖、侵袭和迁移（P<0.01）。用50或60μmol·L厚朴酚处理后，SK-BR-3细胞的凋亡率升高（均P<0.01）；PCNA表达明显下调（P<0.01）。凋亡相关蛋白Bcl-xl和半胱天冬酶-3的细胞内积累减少，但CC3增加。我们还发现与侵袭和迁移相关的蛋白MMP-2表达下调（P<0.01），以及Wnt/β-连环蛋白信号通路相关蛋白β-连环蛋白和c-Myc的表达水平降低（P<0.01）。
厚朴酚可促进SK-BR-3细胞凋亡，并可抑制人乳腺癌SK-BR-3细胞的增殖、迁移和侵袭。其潜在机制可能是通过抑制Wnt信号通路的激活。
