Department of Oncology, The Fifth Hospital of Wuhan, Wuhan, Hubei 430050, P.R. China.
Department of Anesthesiology, The Fifth Hospital of Wuhan, Wuhan, Hubei 430050, P.R. China.
Mol Med Rep. 2020 Jul;22(1):43-50. doi: 10.3892/mmr.2020.11122. Epub 2020 May 5.
Dehydrocorydaline (DHC) is an alkaloid isolated from Corydali syanhusuo that exhibits antitumor properties. It has been reported that DHC can inhibit the proliferation of breast cancer cells, however the underlying molecular mechanism remains elusive. Therefore, the main objective of this study was to evaluate the antitumor activity of DHC, and gain further insights into its mechanism of action. The viability of MDA‑MB‑231 cells was determined through a Cell Counting Kit‑8 assay. The effect of DHC on the proliferation of MDA‑MB‑231 cells was detected by flow cytometry and 5‑ethynyl‑2'‑deoxyuridine staining. Apoptosis was evaluated by Annexin V‑FITC and PI staining through flow cytometry. The impact of DHC treatment on the colony‑forming ability of breast cancer cells was assessed. The expression levels of proliferation‑associated genes cyclin‑dependent kinases 1 (CDK1) and cyclin D1 (CCND1) and apoptosis‑related genes BCL2 and caspases 3/8/9 were quantified by real‑time PCR. Western blot analysis was performed to evaluate the production of cleaved caspase 3/9 and matrix metalloproteinase (MMP)2/9. DHC‑treated MDA‑MB‑231 cells were subcutaneously injected into mice. Subsequent immunohistochemical analyses were performed. DHC inhibited the viability, proliferation, colony‑forming ability and migration of MDA‑MB‑231 cells; in addition, DHC treatment promoted their apoptosis. DHC inhibited the production of proliferation‑ and anti‑apoptosis‑associated proteins CDK1, CCND1, BCL2 as well as that of the metastasis‑associated proteins MMP2 and MMP9. However, it promoted the expression of the pro‑apoptotic caspases 3/8/9. Moreover, DHC inhibited the growth of MDA‑MB‑231 tumor xenografts in SCID mice, and decreased cell proliferation in newly formed tumors in vivo. DHC exerted anticancer effects by downregulating cell proliferation, antiapoptosis, metastasis‑associated proteins CDK1, CCND1, BCL2 and metastasis‑associated proteins MMP2 and MMP9, and by upregulating the expression of proapoptotic proteins caspase 3/8/9.
脱氢紫堇碱(DHC)是从紫堇属植物中分离得到的一种生物碱,具有抗肿瘤特性。据报道,DHC 可抑制乳腺癌细胞的增殖,但作用机制尚不清楚。因此,本研究的主要目的是评估 DHC 的抗肿瘤活性,并进一步探讨其作用机制。通过细胞计数试剂盒-8 测定 MDA-MB-231 细胞的活力。通过流式细胞术和 5-乙炔基-2'-脱氧尿苷染色检测 DHC 对 MDA-MB-231 细胞增殖的影响。通过流式细胞术用 Annexin V-FITC 和 PI 染色评估细胞凋亡。通过集落形成实验评估 DHC 处理对乳腺癌细胞集落形成能力的影响。通过实时 PCR 定量检测增殖相关基因细胞周期蛋白依赖性激酶 1(CDK1)和细胞周期蛋白 D1(CCND1)以及凋亡相关基因 BCL2 和半胱天冬酶 3/8/9 的表达水平。通过 Western blot 分析评估 cleaved caspase 3/9 和基质金属蛋白酶(MMP)2/9 的产生。将 DHC 处理的 MDA-MB-231 细胞皮下注射到小鼠体内。随后进行免疫组织化学分析。DHC 抑制 MDA-MB-231 细胞的活力、增殖、集落形成能力和迁移;此外,DHC 处理促进了细胞凋亡。DHC 抑制增殖和抗凋亡相关蛋白 CDK1、CCND1、BCL2 的产生以及转移相关蛋白 MMP2 和 MMP9 的产生。然而,它促进了促凋亡的半胱天冬酶 3/8/9 的表达。此外,DHC 抑制了 SCID 小鼠中 MDA-MB-231 肿瘤异种移植物的生长,并减少了体内新形成肿瘤中的细胞增殖。DHC 通过下调细胞增殖、抗凋亡、转移相关蛋白 CDK1、CCND1、BCL2 和转移相关蛋白 MMP2 和 MMP9 的表达以及上调促凋亡蛋白半胱天冬酶 3/8/9 的表达发挥抗癌作用。