金花茶调控赖氨酸羟化酶2抑制非小细胞肺癌进展的作用机制研究

Collagen is the key component of extracellular matrix (ECM). Recently, increasing researches showed that the fascicular collagen is relative to the poor prognosis of multiple cancers. PLOD2 catalysis the hydroxylation of collagen lysine residues as a post-translational event in collagen biosynthesis，and the latter forms hydroxylation aldehyde-derived collagen cross-links (HLCCs) which is much more stable. Therefore, PLOD2 is confirmed as the key hydroxylase for stable collagen cross-links. However, as our previous review about PLOD2 in cancer research, there are precious few reports about PLOD2 for lung cancer. And our published research suggested that PLOD2 could not only induce collagen reorganization indirectly promoting NSCLC metastasis, but also directly promote migration and long-term proliferation, to facilitate the poor progression of NSCLC. Nevertheless, the mechanism of PLOD2 promoting progression is still unknown. Meanwhile, our abundant published reports had shown that Camellia nitidissima Chi could inhibit multiple cancers, while their specific mechanisms were also still unknown. Then, in our exploration, we found that Camellia nitidissima Chi could significantly suppress the proliferation and metastasis and inhibit the function and expression of PLOD2 in NSCLC. In this study, we plan to explore anti-cancer effect and mechanism at the molecular, cellular, animal and clinical levels. It will confirm the effect and mechanism of PLOD2 during the NSCLC progression, provide theoretical foundation of PLOD2 as the potential prognostic indicator and reveal a novel therapeutic target for NSCLC. Importantly, this research about the anti-cancer effect of Camellia nitidissima Chi on NSCLC via PLOD2 may provide the scientific proof of Camellia nitidissima Chi for cancer treatment.

胶原是细胞外基质最主要成分，大量研究证实束状排列的胶原与多种肿瘤的不良预后密切相关。其中，PLOD2特异性地羟基化前胶原端肽的赖氨酸，从而形成更稳定的共价交联，成为影响胶原构型的关键酶。我们前期调研发现，针对肺癌，PLOD2涉及的报道极少；并初步证实PLOD2不仅可以调控胶原重构，还可以直接调控肺癌细胞的本身运动能力和体内长期生长能力，但具体作用机制尚未明确。同时，我们实验证实金花茶具有抑制多种肿瘤的作用，但其具体作用机制，亟待揭示。近期，我们又探索发现金花茶可显著抑制NSCLC的增殖和转移，还对PLOD2的功能和表达有抑制作用。因此，本课题拟在分子、细胞、整体动物及临床样本水平，探讨PLOD2在NSCLC病理进展中作用及机制，为其作为潜在预后指标提供充足的理论支撑，揭示治疗肿瘤的新靶点；并开展金花茶通过PLOD2抑制NSCLC进展的作用机制研究，为金花茶用于肿瘤治疗提供充分的科学依据。

胶原是细胞外基质最主要成分，大量研究证实束状排列的胶原与多种肿瘤的不良预后密切相关。其中，PLOD2特异性地羟基化前胶原端肽的赖氨酸，从而形成更稳定的共价交联，成为影响胶原构型的关键酶。同时，我们探索发现金花茶可显著抑制NSCLC的增殖和转移，还对PLOD2的功能和表达有抑制作用。因此，本课题在分子、细胞、整体动物及临床样本水平，探讨PLOD2在NSCLC病理进展中作用及机制，并开展金花茶通过PLOD2抑制NSCLC进展的作用机制研究。研究结果：（1）证实PLOD2在肺癌组织中高表达，与患者不良预后密切相关。（2）证实PLOD2不仅可以调控胶原重构，还可以直接调控肺癌细胞的本身运动能力和体内长期生长能力。（3）基于转录组学揭示，PLOD2通过VEGF、Ras和EGFR等通路发挥调控肿瘤的进展。（4）体内外系列研究证实了金花茶不同有效部位均有一定抗肿瘤作用，尤其是金花茶叶子皂苷部位。通过LC-MS技术对金花茶叶子皂苷部位定性，共鉴定35种化合物，其中25种为皂苷类化合物。（5）金花茶总皂苷可以抑制PLOD2的表达（mRNA和蛋白）和酶学功能（Ⅰ型胶原的排列）。（6）基于转录组学、细胞分子生物学和动物实验揭示，金花茶叶子皂苷可以通过EGFR、 PI3Ks、TGF-β、Trk信号通路等发挥抑制非小细胞肺癌进展作用。总之，本课题为PLOD2作为潜在预后指标提供充足的理论支撑，揭示治疗肿瘤的新靶点；并开展了金花茶通过PLOD2抑制NSCLC进展的作用机制研究，为金花茶用于肿瘤治疗提供充分的科学依据。通过本项目的实施，项目负责人入选湖北省高层次人才计划、中华中医药学会青年人才托举工程等；已发表基金标注论文7篇，其中SCI论文5篇（Q1区2篇,Q2区3篇），中文核心期刊论文2篇（均为T1期刊）；主编英文著作1部(e-book)；学术会议汇报交流2次；培养学生3人，2人获优秀学士论文。