Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, Institute for Clean Energy and Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, PR China.
Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, 400715, PR China.
Lab Chip. 2023 Jun 13;23(12):2838-2853. doi: 10.1039/d3lc00106g.
Breast cancer metastasis to bone is a leading killer in breast cancer patients. A type I collagen-modified thread (Col-I@thread) was prepared for 3-dimensional cell culture and breast cancer bone metastasis co-culture device assembly. First, the coating of Col-I on nylon threads for promoting cell adhesion and growth was studied. Through SEM, XPS, and protein concentration measurements, it was found that the lyophilization method remarkably preserved the Col-I activity and the internal structure of the thread, thereby promoting cell attachment and proliferation. RNA-sequencing (RNA-Seq) and quantitative PCR analysis showed that osteoblast cells (MC3T3-E1) grown on Col-I@thread had elevated RUNX2, ALP, OPN, and Col-I gene expression to promote osteoblast differentiation. Single-cell analysis found that osteoblast MC3T3-E1 cells growing on Col-I@thread had higher Ca secretion activity and mineralized nodules, suggesting robust cell activity and bone matrix formation than cells growing on 2D culture plates. Col-I@threads were knotted in an interdigital cross-finger frame to assemble the breast cancer-bone co-culture model. Confocal microscopy and flow cytometry tests quantified the invasive breast cancer cells. Moreover, the thread-based co-culture devices allowed us to isolate the invasive and non-invasive breast cancer cells to compare their molecular characteristics. qPCR results showed that expression of CX43, CXCR5, and CSPG4 genes was significantly increased in breast cancer cells with bone metastasis. Meanwhile, the expression of RUNX2 and OPG genes in osteoblasts was inhibited. The co-culture model based on the Col-I@thread mimics the bone tissue microenvironment to reveal the cross-talk between cancer cells and bone tissue. Moreover, the thread-based co-culture device is easy to fabricate and operate, providing a platform for exploring the cellular and molecular mechanisms of breast cancer bone metastasis, and holds potential for high-throughput screening of anti-breast cancer bone metastasis drugs.
乳腺癌骨转移是乳腺癌患者的主要致死原因。我们制备了一种 I 型胶原蛋白修饰线（Col-I@thread），用于三维细胞培养和乳腺癌骨转移共培养装置的组装。首先，我们研究了 Col-I 在尼龙线上的涂层，以促进细胞黏附和生长。通过 SEM、XPS 和蛋白质浓度测量，我们发现冻干法能显著保持 Col-I 的活性和线的内部结构，从而促进细胞附着和增殖。RNA 测序（RNA-Seq）和定量 PCR 分析表明，在 Col-I@thread 上生长的成骨细胞（MC3T3-E1）的 RUNX2、ALP、OPN 和 Col-I 基因表达升高，促进成骨细胞分化。单细胞分析发现，在 Col-I@thread 上生长的成骨细胞 MC3T3-E1 具有更高的 Ca 分泌活性和矿化结节，表明其细胞活性和骨基质形成能力强于在 2D 培养板上生长的细胞。我们将 Col-I@thread 打结在指形交叉框架中，以组装乳腺癌-骨共培养模型。共聚焦显微镜和流式细胞术测试定量了侵袭性乳腺癌细胞。此外，基于线的共培养装置允许我们分离侵袭性和非侵袭性乳腺癌细胞，以比较它们的分子特征。qPCR 结果表明，具有骨转移的乳腺癌细胞中 CX43、CXCR5 和 CSPG4 基因的表达显著增加，同时成骨细胞中 RUNX2 和 OPG 基因的表达受到抑制。基于 Col-I@thread 的共培养模型模拟了骨组织微环境，揭示了癌细胞与骨组织之间的相互作用。此外，基于线的共培养装置易于制造和操作，为探索乳腺癌骨转移的细胞和分子机制提供了平台，并且具有用于高通量筛选抗乳腺癌骨转移药物的潜力。
