Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, 46556, USA.
Harper Cancer Research Institute, University of Notre Dame, South Bend, IN, 46617, USA.
Sci Rep. 2017 Nov 10;7(1):15295. doi: 10.1038/s41598-017-15547-7.
Ovarian cancer (OvCa) cells are reported to undergo biochemical changes at the cell surface in response to treatment with lysophosphatidic acid (LPA). Here we use scanning electron microscopy (SEM) and multiplex coherent anti-Stokes Raman scattering (CARS) imaging via supercontinuum excitation to probe morphological changes that result from LPA treatment. SEM images show distinct shedding of microvilli-like features upon treatment with LPA. Analysis of multiplex CARS images can distinguish between molecular components, such as lipids and proteins. Our results indicate that OvCa429 and SKOV3ip epithelial ovarian cancer cells undergo similar morphological and chemical responses to treatment with LPA. The microvilli-like structures on the surface of multicellular aggregates (MCAs) are removed by treatment with LPA. The CARS analysis shows a distinct decrease in protein and increase in lipid composition on the surface of LPA-treated cells. Importantly, the CARS signals from cellular sheddings from MCAs with LPA treatment are consistent with cleavage of proteins originally present. Mass spectrometry on the cellular sheddings show that a large number of proteins, both membrane and intracellular, are present. An increased number of peptides are detected for the mesenchymal cell line relative to the epithelial cell indicating a differential response to LPA treatment with cancer progression.
据报道,卵巢癌细胞在受到溶血磷脂酸 (LPA) 治疗时,其细胞表面会发生生化变化。在这里,我们使用扫描电子显微镜 (SEM) 和基于超连续谱激发的多路相干反斯托克斯拉曼散射 (CARS) 成像来探测由 LPA 处理引起的形态变化。SEM 图像显示,在用 LPA 处理后,微绒毛样特征明显脱落。对多路 CARS 图像的分析可以区分脂质和蛋白质等分子成分。我们的结果表明,上皮性卵巢癌细胞 OvCa429 和 SKOV3ip 对 LPA 的治疗表现出相似的形态和化学反应。多细胞聚集体 (MCA) 表面的微绒毛样结构在用 LPA 处理后被去除。CARS 分析表明,在用 LPA 处理的细胞表面,蛋白质组成明显减少,脂质组成增加。重要的是,用 LPA 处理 MCA 后细胞脱落的 CARS 信号与原有的蛋白质切割一致。对细胞脱落物进行质谱分析表明,存在大量的蛋白质,包括膜蛋白和胞内蛋白。与上皮细胞相比,间充质细胞系检测到的肽数量增加,表明在癌症进展过程中对 LPA 治疗有不同的反应。