Abstract:
Experiments conducted on triple-negative breast cancer have shown that fucoidan from Lessonia trabeculata (FLt) exhibits cytotoxic and antitumor properties. However, further research is necessary to gain a complete understanding of its bioactivity and level of cytotoxicity. The cytotoxic effect of FLt was determined by the 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Apoptosis was analyzed using annexin V and caspase 3/7 staining kit and DNA fragmentation. In addition, transcriptional expression of antiapoptotic (Bcl-2 and XIAP) and proapoptotic (caspase 8, caspase 9, and AIF) genes were analyzed in TNBC 4T1 cells. After 72 h of culture, the IC50 for FLt was 561 μg/mL, while doxorubicin (Dox) had an IC50 of 0.04 μg/mL. In addition, assays for FLt + Dox were performed. Annexin V and caspase 3/7 revealed that FLt induces early and late-stage apoptosis. DNA fragmentation results support necrotic death of 4T1 cells. Similarly, transcripts that prevent cell death were decreased, while transcripts that promote cell death were increased. This study showed that FLt induces apoptosis by both caspase-dependent and caspase-independent mechanisms. These findings suggest that FLt may have potential applications in breast cancer treatment. Further research will provide more information to elucidate the mechanism of action of FLt.
摘要:
在三阴性乳腺癌上进行的实验表明,来自小梁的岩藻依聚糖(FLt)表现出细胞毒性和抗肿瘤特性。然而,需要进一步的研究才能全面了解其生物活性和细胞毒性水平。FLt的细胞毒性作用通过2,5-二苯基-2H-四唑溴化物(MTT)测定来确定。使用膜联蛋白V和半胱天冬酶3/7染色试剂盒和DNA片段化分析细胞凋亡。此外,在TNBC4T1细胞中分析了抗凋亡(Bcl-2和XIAP)和促凋亡(caspase8,caspase9和AIF)基因的转录表达。培养72小时后,FLt的IC50为561μg/mL,而多柔比星(Dox)的IC50为0.04μg/mL。此外,进行FLt+Dox的测定。膜联蛋白V和caspase3/7显示FLt诱导早期和晚期细胞凋亡。DNA片段化结果支持4T1细胞的坏死死亡。同样,防止细胞死亡的转录物减少了,而促进细胞死亡的转录物增加。这项研究表明,FLt通过caspase依赖性和caspase非依赖性机制诱导细胞凋亡。这些发现表明FLt可能在乳腺癌治疗中具有潜在的应用。进一步的研究将为阐明FLt的作用机制提供更多信息。