JASPLAKINOLIDES INHIBITION OF THE GROWTH OF PROSTATE CARCINOMA-CELLS IN-VITRO WITH DISRUPTION OF THE ACTIN CYTOSKELETON
JASPLAKINOLIDES INHIBITION OF THE GROWTH OF PROSTATE CARCINOMA-CELLS IN-VITRO WITH DISRUPTION OF THE ACTIN CYTOSKELETON
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DOI：
10.1093/jnci/87.1.46
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发表时间：
1995-01-04
期刊：
影响因子：
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通讯作者：
DUNCAN, KLK
中科院分区：
文献类型：
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作者：
SENDEROWICZ, AMJ;KAUR, G;DUNCAN, KLK
背景：Jasplakinolide 是一种由印度太平洋海绵 Jaspis johnstoni 产生的环缩酚肽，据报道可以抑制乳腺癌细胞的生长。目的：研究了茉莉酮内酯对三种人永生化前列腺癌细胞系（PC-3、LNCaP 和 TSU-Pr1）增殖的影响。详细研究了 jasplakinolide 对 PC-3 细胞系的生长抑制作用，以阐明其作用机制。方法：细胞计数仅用于研究生长抑制，基于蛋白质的微孔板测定用于评估引起持续生长抑制所需的暴露时间并研究茉莉酮内酯类似物的作用。通过掺入放射性标记前体来评估代谢变化。使用罗丹明鬼笔环肽 (RP) 和细胞骨架成分抗体，通过荧光显微镜研究了茉莉酮内酯对细胞骨架的影响。通过从固定细胞中提取结合的荧光物质并在分光荧光计中测量荧光量来量化 RP 结合的变化。结果：Jasplakinolide 作用 48 小时，可有效抑制 PC-3、LNCaP 和 TSU-Pr1 细胞的生长；导致50%生长抑制的茉莉酮内酯剂量对于PC-3细胞为65nM，对于LNCaP细胞为41nM，对于TSU-Pr1细胞为170nhl。在 PC-3 细胞中，暴露于 160 nM 48 小时会导致完全生长抑制，即使在药物去除后，这种抑制仍持续数天。几种茉莉酮内酯类似物也抑制PC-3细胞的生长，尽管改变大环内酯环刚性的类似物无效。暴露于茉莉酮内酯后，PC-3 细胞中的 DNA、RNA 或蛋白质合成或细胞内三磷酸腺苷水平没有出现早期变化。茉莉花内酯的生长抑制伴随着显着的形态变化。暴露几次倍增导致多核细胞。对 PC-3 细胞中这些变化的进一步研究揭示了肌动蛋白细胞骨架的显着且早期破坏以及 RP 结合的统计显着减少。茉莉酮内酯的剂量、暴露时间以及结构类似物的生长抑制模式与肌动蛋白分布的变化相对应。结论： Jasplakinolide 是一种新型海洋天然产物，具有针对人前列腺癌细胞系的专利体外抗增殖活性，并且它似乎以肌动蛋白细胞骨架为目标。意义： Jasplakinolide 是进一步临床前开发的潜在候选者，也是一类可以破坏哺乳动物细胞中肌动蛋白细胞骨架的新型治疗剂的先导结构。
Background: Jasplakinolide, a cyclodepsipeptide produced by an Indo-Pacific sponge, Jaspis johnstoni, has been reported to inhibit the growth of breast cancer cells. Purpose: The effects of jasplakinolide on the proliferation of three human immortalized prostate carcinoma cell lines (PC-3, LNCaP, and TSU-Pr1) were studied. The growth-inhibitory effect of jasplakinolide on the PC-3 cell line was studied in detail to elucidate its mechanism of action. Methods: Cell counts mere used to study growth inhibition, a protein-based microplate assay was used to assess the time of exposure needed to cause persistent growth inhibition and to study the effects of jasplakinolide analogues. Metabolic changes were assessed by following the incorporation of radiolabeled precursors. The effects of jasplakinolide on the cytoskeleton were studied by fluorescent microscopy, using rhodamine phalloidin (RP) and antibodies to cytoskeletal components. Changes in RP binding were quantified by extracting bound fluorescent material from fixed cells and measuring the amount of fluorescence in a spectrofluorometer. Results: The growth of PC-3, LNCaP, and TSU-Pr1 cells was potently inhibited by exposure to jasplakinolide for 48 hours; doses of jasplakinolide that led to 50% growth inhibition were 65 nM for PC-3 cells, 41 nM for LNCaP cells, and 170 nhl for TSU-Pr1 cells. In PC-3 cells, exposure to 160 nM for 48 hours led to total growth inhibition, which persisted for several days even after drug removal. Several jasplakinolide analogues also inhibited the growth of PC-3 cells, although analogues in which the rigidity of the macrolide ring was altered were ineffective. No early changes in the synthesis of DNA, RNA, or protein or in intracellular adenosine triphosphate levels were seen in the PC-3 cells after exposure to jasplakinolide. Growth inhibition by jasplakinolide was accompanied by striking morphologic changes. Exposure for several doublings led to multinucleated cells. Further investigation of these changes in the PC-3 cells revealed a dramatic and early disruption of the actin cytoskeleton and a statistically significant decrease in RP binding. The doses of jasplakinolide, the time of exposure, and the pattern of growth inhibition by structural analogues corresponded with the changes seen in actin distribution. Conclusions: Jasplakinolide represents a novel marine natural product with patent in vitro antiproliferative activity against human prostate carcinoma cell lines, and it appears to target the actin cytoskeleton. Implications: Jasplakinolide is a potential candidate for further preclinical development and a lead structure for a novel class of therapeutic agents that can disrupt the actin cytoskeleton in mammalian cells.