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黑寡妇幼蛛提取物对HeLa细胞的细胞毒性和凋亡活性。

Cytotoxic and apoptotic activities of black widow spiderling extract against HeLa cells.

作者信息

Peng Xiaozhen, Dai Zhipan, Lei Qian, Liang Long, Yan Shuai, Wang Xianchun

机构信息

Key Laboratory of Protein Chemistry and Developmental Biology of the Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, P.R. China.

Department of Cell Biology, School of Life Sciences, Central South University, Changsha, Hunan 410013, P.R. China.

出版信息

Exp Ther Med. 2017 Jun;13(6):3267-3274. doi: 10.3892/etm.2017.4391. Epub 2017 Apr 27.

Abstract

Black widow spiders contain toxic components not only in the venom glands but also in other parts of the spider body, including the legs and abdomen. Additionally, both the eggs and newborn spiderlings of the black widow spider contain venom. It is important to investigate their potential effects on cancer cells. In the present study, the effects of newborn black widow spiderling extract on human HeLa cells were evaluated . When applied at different concentrations, the total extract decreased HeLa cell viability in a dose-dependent manner, with an IC value of 158 µg/ml. Flow cytometry indicated that treatment of HeLa cells with the total extract of the spiderlings induced apoptosis in HeLa cells in a dose-dependent manner and led to cell cycle arrest in the S-phase. Additionally, application of the total extract at different concentrations increased apoptosis-related caspase 3 activity in a dose-dependent manner. HeLa cells treated with the total extract appeared to be morphologically changed, exhibiting membrane blebbing, nuclear fragmentation and condensation of chromatin. Further separation and activity screening demonstrated that the cytotoxic and apoptotic activities of the total extract were attributable mainly to its high molecular mass proteins, one of which was purified and characterized to determine its anti-tumor activities on HeLa cells. The results of the present study therefore have expanded understanding regarding the effect of spider toxins on cancer cells and suggested that components of black widow spiderlings may be developed as a promising novel agent to treat cancer.

摘要

黑寡妇蜘蛛不仅在毒腺中含有有毒成分,在蜘蛛身体的其他部位,包括腿部和腹部也含有有毒成分。此外,黑寡妇蜘蛛的卵和新生幼蛛都含有毒液。研究它们对癌细胞的潜在影响很重要。在本研究中,评估了新生黑寡妇幼蛛提取物对人宫颈癌细胞(HeLa细胞)的影响。当以不同浓度应用时,总提取物以剂量依赖的方式降低了HeLa细胞的活力,IC值为158微克/毫升。流式细胞术表明,用幼蛛总提取物处理HeLa细胞会以剂量依赖的方式诱导HeLa细胞凋亡,并导致细胞周期停滞在S期。此外,以不同浓度应用总提取物会以剂量依赖的方式增加凋亡相关的半胱天冬酶3活性。用总提取物处理的HeLa细胞在形态上似乎发生了变化,表现出细胞膜泡化、核碎片化和染色质凝聚。进一步的分离和活性筛选表明,总提取物的细胞毒性和凋亡活性主要归因于其高分子量蛋白质,其中一种蛋白质被纯化并进行了表征,以确定其对HeLa细胞的抗肿瘤活性。因此,本研究结果扩展了对蜘蛛毒素对癌细胞影响的认识,并表明黑寡妇幼蛛的成分可能被开发成为一种有前景的新型抗癌药物。

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