Institute of virology and AIDS research, The first hospital of Jilin University, Changchun 130021, PR China.
Int J Med Sci. 2013 Jul 7;10(9):1149-56. doi: 10.7150/ijms.6275. Print 2013.
Dracorhodin perchlorate (DP) was a synthetic analogue of the antimicrobial anthocyanin red pigment dracorhodin. It was reported that DP could induce apoptosis in human prostate cancer, human gastric tumor cells and human melanoma, but the cytotoxic effect of DP on human breast cancer was not investigated. This study would investigate whether DP was a candidate chemical of anti-human breast cancer.
The MTT assay reflected the number of viable cells through measuring the activity of cellular enzymes. Phase contrast microscopy visualized cell morphology. Fluorescence microscopy detected nuclear fragmentation after Hoechst 33258 staining. Flowcytometric analysis of Annexin V-PI staining and Rodamine 123 staining was used to detect cell apoptosis and mitochondrial membrane potential (MMP). Real time PCR detected mRNA level. Western blot examined protein expression.
DP dose and time-dependently inhibited the growth of MCF-7 cells. DP inhibited MCF-7 cell growth through apoptosis. DP regulated the expression of Bcl-2 and Bax, which were mitochondrial pathway proteins, to decrease MMP, and DP promoted the transcription of Bax and inhibited Bcl-2. Apoptosis-inducing factor (AIF) and cytochrome c which localized in mitochondrial in physiological condition were released into cytoplasm when MMP was decreased. DP activated caspase-9, which was the downstream of mitochondrial pathway. Therefore DP decreased MMP to release AIF and cytochrome c into cytoplasm, further activating caspase 9, lastly led to apoptosis.
Therefore DP was a candidate for anti-breast cancer, DP induced apoptosis of MCF-7 through mitochondrial pathway.
过氯酸血根碱（DP）是一种具有抗菌作用的花色苷红色素血根碱的合成类似物。有报道称 DP 可诱导人前列腺癌、人胃癌细胞和人黑色素瘤细胞凋亡，但 DP 对人乳腺癌的细胞毒性作用尚未得到研究。本研究旨在探讨 DP 是否是一种抗人乳腺癌的候选化学物质。
MTT 检测通过测量细胞酶的活性来反映活细胞的数量。相差显微镜观察细胞形态。Hoechst 33258 染色后荧光显微镜检测核碎片。用 Annexin V-PI 染色和罗丹明 123 染色的流式细胞术分析检测细胞凋亡和线粒体膜电位（MMP）。实时 PCR 检测 mRNA 水平。Western blot 检测蛋白表达。
DP 呈剂量和时间依赖性抑制 MCF-7 细胞的生长。DP 通过凋亡抑制 MCF-7 细胞生长。DP 调节线粒体途径蛋白 Bcl-2 和 Bax 的表达，降低 MMP，DP 促进 Bax 的转录并抑制 Bcl-2。生理条件下位于线粒体中的凋亡诱导因子（AIF）和细胞色素 c 在 MMP 降低时被释放到细胞质中。DP 激活了 caspase-9，这是线粒体途径的下游。因此，DP 通过线粒体途径降低 MMP，释放 AIF 和细胞色素 c 进入细胞质，进一步激活 caspase 9，最终导致细胞凋亡。
因此 DP 是一种抗乳腺癌的候选药物，DP 通过线粒体途径诱导 MCF-7 细胞凋亡。
