格里菲帕黄酮诱导人乳腺癌 MCF-7 细胞体外凋亡。

Griffipavixanthone induces apoptosis of human breast cancer MCF-7 cells in vitro.

机构信息

Shanxi Key Laboratory of Carcinogenesis and Translational Research of Esophageal Cancer, Shanxi Medical University, Taiyuan, Shanxi, 030001, People's Republic of China.

Key Laboratory of Cellular Physiology, Ministry of Education, Shanxi Medical University, Taiyuan, Shanxi, 030001, People's Republic of China.

出版信息

Breast Cancer. 2019 Mar;26(2):190-197. doi: 10.1007/s12282-018-0912-2. Epub 2018 Sep 26.

DOI:10.1007/s12282-018-0912-2

PMID:30259331

Abstract

BACKGROUND

Griffipavixanthone (GPX) is a compound extracted from Garcinia oblongifolia Champ. But, no research has yet been done about the effect of GPX on breast cancer.

METHODS

We evaluated the proliferation of human breast cancer cells by CCK-8 assay and apoptosis by Annexin V (AV)-FITC and PI double staining. We used transwell assay to indicate the invasion and migration of MCF-7. To explore the molecular mechanism of GPX, we detected the mRNA and protein expression using qRT-PCR and Western blot.

RESULTS

In this study, we evaluated if GPX could inhibit the proliferation of human breast cancer cell MCF-7 and T-47D with IC50 value of 9.64 ± 0.12 µM and 10.2 1 ± 0.38 µM at 48 h. And the IC50 value of MCF-10A is 32.11 ± 0.21 µM, which showed GPX had a tiny side effect for normal breast cells. Annexin V (AV)-FITC and PI double staining demonstrated firmly the apoptosis of MCF-7 resulting from GPX. Transwell assay indicated that GPX inhibited the invasion and migration of MCF-7. In addition, we found GPX cleaved caspase-8/9 and PARP, which play important roles in apoptotic pathway. Furthermore, through the Western blot assay, GPX increased the level of pro-apoptosis protein Bax and cytochrome C. On the contrary, GPX decreased the level of anti-apoptosis protein Bcl-2. Moreover, GPX increased the mRNA and protein expression level of p53 and its target genes, which indicated that GPX induced MCF-7 cell apoptosis by up-regulating p53 and Bax expression while suppressing Bcl-2 expression.

CONCLUSION

All the results showed that GPX induces MCF-7 cell apoptosis and could be considered as a potential drug for breast cancer.

摘要

背景

Griffipavixanthone (GPX) 是从藤黄属植物中提取的一种化合物。但是，目前还没有关于 GPX 对乳腺癌影响的研究。

方法

我们通过 CCK-8 法评估了人乳腺癌细胞的增殖，通过 Annexin V (AV)-FITC 和 PI 双重染色评估了细胞凋亡。我们使用 Transwell 测定法来指示 MCF-7 的侵袭和迁移。为了探讨 GPX 的分子机制，我们使用 qRT-PCR 和 Western blot 检测了 mRNA 和蛋白表达。

结果

在这项研究中，我们评估了 GPX 是否能抑制人乳腺癌细胞 MCF-7 和 T-47D 的增殖，在 48 小时时，其 IC50 值分别为 9.64±0.12μM 和 10.21±0.38μM。而 MCF-10A 的 IC50 值为 32.11±0.21μM，这表明 GPX 对正常乳腺细胞有很小的副作用。Annexin V (AV)-FITC 和 PI 双重染色证实了 GPX 诱导 MCF-7 细胞凋亡。Transwell 测定表明，GPX 抑制了 MCF-7 的侵袭和迁移。此外，我们发现 GPX 切割了 caspase-8/9 和 PARP，它们在凋亡途径中起着重要作用。此外，通过 Western blot 测定，GPX 增加了促凋亡蛋白 Bax 和细胞色素 C 的水平。相反，GPX 降低了抗凋亡蛋白 Bcl-2 的水平。此外，GPX 增加了 p53 及其靶基因的 mRNA 和蛋白表达水平，这表明 GPX 通过上调 p53 和 Bax 的表达，同时抑制 Bcl-2 的表达，诱导 MCF-7 细胞凋亡。