Polarization-resolved second harmonic generation imaging of human ovarian cancer.
Polarization-resolved second harmonic generation imaging of human ovarian cancer.
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DOI：
10.1117/1.jbo.23.6.066501
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发表时间：
2018-06
影响因子：
3.5
通讯作者：
Campagnola PJ
中科院分区：
文献类型：
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作者：
Campbell KR;Chaudhary R;Handel JM;Patankar MS;Campagnola PJ
人卵巢癌细胞外基质的重塑可表现为胶原浓度升高、原纤维/纤维内排列改变和/或不同胶原亚型的上调。我们使用基于像素的二次谐波产生（SHG）偏振显微镜分析来探测人类卵巢组织[正常间质，良性肿瘤和高级别浆液（HGS）肿瘤]的分子变化：(i)通过单轴分子模型确定-螺旋螺距角，（ii）通过SHG各向异性确定原纤维内的胶原排列，以及（iii）通过SHG圆二色性（SHG- cd）确定手性。由于矩阵的复杂结构缺乏高度的纤维对齐，因此需要像素方法。正常间质和良性肿瘤的螺旋节角差异最大，这与Col III亚型在良性肿瘤中上调的基因表达一致。这些数据与之前报道的HGS肿瘤中Col III的上调不一致。不同组织也表现出不同的SHG各向异性和SHG- cd反应，这与良性和恶性肿瘤中Col III的掺入或Col I序列的随机化一致。此外，高级别肿瘤显示胶原蛋白浓度较高，这种结缔组织形成与这些组织中较高的纤维密度一致。这些结果共同表明，所有组织中的原纤维组合都是不同的，这些差异可能是由于胶原蛋白的合成而不是现有胶原蛋白的重塑。重要的是，这些分析是无标记的，并询问完整组织的亚分辨率胶原结构，而不需要传统的结构生物学工具。
Remodeling of the extracellular matrix in human ovarian cancer can be manifested in increased collagen concentration, changes in alignment within fibrils/fibers and/or up-regulation of different collagen isoforms. We used pixel-based second harmonic generation (SHG) polarization microscopy analyses to probe these molecular changes in human ovarian tissues [normal stroma, benign tumors, and high-grade serous (HGS) tumors] by: (i) determination of the -helical pitch angle via the single-axis molecular model, (ii) collagen alignment within fibrils via SHG anisotropy, and (iii) chirality via SHG circular dichroism (SHG-CD). Pixel approaches are required due to the complex structure of the matrix that lacks a high degree of fiber alignment. The largest differences in the helical pitch angle were between normal stroma and benign tumors, consistent with gene expression showing the Col III isoform is up-regulated in the latter. The data were not consistent with up-regulation of Col III in HGS tumors as previous reports have suggested. The different tissues also displayed differing SHG anisotropies and SHG-CD responses, consistent with either Col III incorporation or randomization of Col I alignment within benign and malignant tumors. Additionally, the high-grade tumors displayed higher collagen concentration, where this desmoplasia is consistent with the higher fiber density in these tissues. These results collectively indicate that the fibril assemblies are distinct in all tissues, where these differences likely result from the synthesis of collagen rather than remodeling of existing collagen. Importantly, these analyses are label-free and interrogate subresolution collagen structure on intact tissues, without the need for conventional structural biology tools.