肺癌骨轉移是全球癌症轉移相關頭號死亡病例，腫瘤細胞與其轉移器官微環境之間的交流對於日後的腫瘤轉移形態和腫瘤演化進程是至關重要的。骨骼守恆依賴成骨細胞與蝕骨細胞之間的平衡分化活動來維持，並在腫瘤骨轉移中扮演重要角色。Runt相關轉錄因子2 (Runx2) 蛋白質表現在由老鼠間質幹細胞分化至成熟成骨细胞時呈下調。我們證明高與低趨骨轉移肺癌細胞之培養上清液均可抑制成骨細胞分化並抑制Runx2蛋白表現之下調，而蝕骨細胞的分化和整合素 (integrin) β3蛋白質之表現以及由蝕骨細胞所分泌的組織蛋白酶k在肺癌細胞培養上清液作用下則不受干擾。我們研究結果顯示，肺癌細胞所分泌的分泌小體對成骨細胞分化並沒有影響，雖然肺癌細胞分泌小體的存在也不會影響蝕骨細胞之巨大多核細胞形態，但是高趨骨轉移性肺癌細胞的分泌小體會干擾蝕骨細胞整合素β3膜蛋白表現量以及由蝕骨細胞所分泌的組織蛋白酶k的活性。然而，分泌小體離心上清液仍可抑制成骨細胞分化並抑制Runx2蛋白表現之下調。總結來說，我們的研究結果指出肺癌細胞培養上清液抑制成骨細胞分化源之於分泌小體離心上清液，以及分泌小體可能藉由內含物傳輸機制增強肺癌細胞骨轉移功能，顯示趨骨肺癌細胞分泌物可透過干擾骨骼守恆以及活化腫瘤細胞來促進骨轉移現象。

Osteotropic lung cancer metastases are dominant causes of cancer-associated mortality worldwide. Crosstalk between tumor cells and secondary organ metastatic site microenvironment is critical for metastatic niche formation and tumor progression. Balancing differentiation activity of osteoblasts and osteoclasts is essential for bone homeostasis. Runx2 protein expression is down-regulated during differentiation of murine mesenchymal stem cells into mature osteoblasts. We demonstrated that high and low murine osteotropic lung cancer cell-conditioned medium inhibited osteoblast differentiation and prevented Runx2 protein down -regulation, while integrin β3 membrane protein expression and secreted cathepsin k enzyme activity of osteoclasts was not interrupted by lung cancer cell-conditioned medium. Lung cancer cell-derived exosomes did not inhibit osteoblast differentiation and prevent Runx2 protein down-regulation. Although lung cancer cell-derived exosomes did not affected osteoclast multinucleated cell morphology either, high osteotropic lung cancer cell-derived exosomes did interrupt integrin β3 membrane protein expression and secreted cathepsin k enzyme activity of osteoclasts. However, exosome-depleted supernatant inhibited osteoblast differentiation and prevented Runx2 protein down-regulation. Here, we show that lung cancer cell-derived exosomes have no effect on osteoblast differentiation while high osteotropic lung cancer cell-derived exosomes interrupt osteoclastic activity. In summary, our results demonstrated the inhibitory effect of lung cancer cell-conditioned medium on osteoblast differentiation originates from exosome-depleted supernatants, and suggest that lung cancer cell-derived exosome is able to enhance lung cancer cell osteotropic metastatic behavior through cargo material transferring mechanism. Our results support the notion that osteotropic lung cancer cell-secreted factors facilitate osteotropic metastasis through interrupting bone homeostasis and priming tumor cells.

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