Department of Genetics and Morphology, University of Brasília, Asa Norte, Brazil; Department of Cell Biology, University of Brasília, Asa Norte, Brazil.
Cancer Lett. 2010 Jul 1;293(1):73-81. doi: 10.1016/j.canlet.2009.12.017. Epub 2010 Feb 4.
In this work, we report the effects of a Bowman-Birk protease inhibitor, the Black-Eyed Pea Trypsin/Chymotrypsin Inhibitor - BTCI, purified from Vigna unguiculata seeds, on the MCF-7 breast cancer cells. The treatment of MCF-7 with 200microM BTCI for 72h induced significant reduction of the cell viability and proliferation (arrest in S and G2/M phase). These cytostatic effects were accompanied by acute morphological modifications including the alteration of the nuclear morphology, plasma membrane fragmentation, cytoplasm disorganization, presence of double-membrane vesicles, mitochondrial swelling, and an increase in the size of lysosomes. Significative DNA fragmentation, annexin-V(+) cell number increase, mitochondrial membrane potential reduction, and cytoplasm acidification were also detected. All together, these cytostatic and cytotoxic results point out to BTCI-induced apoptosis cell death associated with severe cell morphological alterations and lysosome membrane permeabilization. Our study confirms the anticarcinogenic potential of Bowman-Birk protease inhibitors and identifies BTCI as a promising tool for drug developments aimed at the treatment of breast cancer.
在这项工作中，我们报告了从 Vigna unguiculata 种子中纯化得到的 Bowman-Birk 蛋白酶抑制剂——黑眼豆胰蛋白酶/糜蛋白酶抑制剂（BTCI）对 MCF-7 乳腺癌细胞的影响。用 200μM BTCI 处理 MCF-7 细胞 72 小时，诱导细胞活力和增殖显著降低（S 和 G2/M 期停滞）。这些细胞生长抑制作用伴随着急性形态学改变，包括核形态改变、质膜碎片化、细胞质紊乱、双层膜囊泡出现、线粒体肿胀和溶酶体增大。还检测到显著的 DNA 片段化、膜联蛋白 V（+）细胞数量增加、线粒体膜电位降低和细胞质酸化。所有这些细胞生长抑制和细胞毒性结果都表明 BTCI 诱导的细胞凋亡与严重的细胞形态改变和溶酶体膜通透性有关。我们的研究证实了 Bowman-Birk 蛋白酶抑制剂的抗癌潜力，并确定 BTCI 是一种有前途的药物开发工具，旨在治疗乳腺癌。
