Sumalatha Kodandaram, Gowda Mohan, Meenakshisundaram Sreepriya
J Complement Integr Med. 2017 Mar 16;14(2). doi: 10.1515/jcim-2016-0131.
Background Embelin is a benzoquinone reported to possess anticancer activity in several in vivo and in vitro models of carcinogenesis, especially hematopoietic and prostate malignancy. A detailed investigation on the influence of embelin on epithelial malignancy model system, especially colon adenocarcinoma, is lacking. The objective of the current study is to investigate the antiproliferative, antiinvasive and proapoptotic potential of embelin on colon adenocarcinoma cell line HT-29. Methods The effect of embelin (35 µg/mL for 24 h) on cell proliferation was assessed by Sulforhodamine B assay and bromodeoxyuridine incorporation test, antiinvasive effect by Boyden chamber assay and scratch assay. Proapoptotic effects of embelin were determined by studies on DNA fragmentation, annexin V-FITC labeling, TUNEL assay, COMET assay and assay of caspase-3 activity. Influence of embelin on the expression of genes regulating apoptosis (caspase 3 and 9, Bcl-2, Bax, cytochrome C and X-linked inhibitor of apoptosis protein) and migration/invasion (matrix metalloproteinase [MMP]-2 and MMP-9) was investigated by reverse transcription polymerase chain reaction (PCR). Further, the effect of embelin on the levels of reactive oxygen species (ROS), lipid peroxides, nitric oxide, mitochondrial membrane potential and antioxidant status (total reduced glutathione [GSH] and GSH-S-transferase) was evaluated. Results Results implicated that embelin treatment inhibited proliferation (IC50 35 µg/mL), induced DNA fragmentation, phosphatidyl serine externalization, increased caspase expression, decreased cell migration and expression of MMPs in HT-29 cells. Interestingly, embelin exhibited prooxidant effect on HT-29 cells and induced excessive ROS generation resulting in apoptotic cell death. Conclusions To conclude, embelin treatment could be a promising strategy for the chemotherapy of colon cancer.
背景 紫铆因是一种苯醌，据报道在多种体内和体外致癌模型中具有抗癌活性，尤其是对造血系统和前列腺恶性肿瘤。目前尚缺乏对紫铆因对上皮恶性肿瘤模型系统，特别是结肠腺癌影响的详细研究。本研究的目的是探讨紫铆因对结肠腺癌细胞系HT-29的抗增殖、抗侵袭和促凋亡潜力。方法 通过磺酰罗丹明B法和溴脱氧尿苷掺入试验评估紫铆因（35 μg/mL，作用24小时）对细胞增殖的影响，通过Boyden小室试验和划痕试验评估其抗侵袭作用。通过DNA片段化研究、膜联蛋白V-FITC标记、TUNEL试验、彗星试验和半胱天冬酶-3活性测定来确定紫铆因的促凋亡作用。通过逆转录聚合酶链反应（PCR）研究紫铆因对调节细胞凋亡（半胱天冬酶3和9、Bcl-2、Bax、细胞色素C和X连锁凋亡抑制蛋白）和迁移/侵袭（基质金属蛋白酶[MMP]-2和MMP-9）相关基因表达的影响。此外，评估了紫铆因对活性氧（ROS）、脂质过氧化物、一氧化氮、线粒体膜电位和抗氧化状态（总还原型谷胱甘肽[GSH]和GSH-S-转移酶）水平的影响。结果 结果表明，紫铆因处理可抑制HT-29细胞的增殖（IC50为35 μg/mL），诱导DNA片段化、磷脂酰丝氨酸外化，增加半胱天冬酶表达，减少细胞迁移和MMPs的表达。有趣的是，紫铆因对HT-29细胞表现出促氧化作用，诱导过量ROS生成，导致凋亡性细胞死亡。结论 总之，紫铆因治疗可能是结肠癌化疗的一种有前景的策略。
