前列腺癌细胞外泌体抑制成骨细胞分化

Prostate cancer exosomes inhibit osteoblast differentition

目的:探讨前列腺癌细胞外泌体对成骨细胞分化的影响。方法:采用ExoQuick-TC试剂盒与超滤法结合分离纯化前列腺癌细胞系PC3的外泌体,通过电镜、外泌体蛋白标志物CD63进行鉴定。用纯化的前列腺癌细胞外泌体处理前成骨细胞,提取总RNA,采用荧光实时定量PCR检测早期成骨分化标志物Runx2、Osx和中期成骨分化标志物Alp的表达情况。此外利用碱性磷酸酶(alkaline phosphatase,ALP)染色、茜素红染色方法检测成骨分化情况。结果:前列腺癌细胞外泌体可以进入到前成骨细胞中,并且外泌体处理前成骨细胞后,与对照组相比,Alp、Runx2与Osx的表达量显著降低(P <0.05),染色结果显示成骨分化受抑制。结论:在前列腺癌中,前列腺癌细胞会分泌出外泌体作用于前成骨细胞,并抑制成骨细胞的分化。

Objective:This study aims to explore the effect of exosome of prostate cancer cells on the osteoblast differentiation.Methods:The ExoQuick-TC kit and ultrafiltration are adopted to isolate and purify exosomes from prostate cancer cell lines.The exosomes will be characterized by marker CD63,and observed through electron microscopy.Extract total RNA from pre-osteoblasts treated by exosomes of prostate cancer cells;then expressions of Osx,Runx2 and Alp were detected by real-time PCR.Osx and Runx2 are the biomarker of early osteogenic differentiation and Alp is the biomarker of metaphase osteogenic differentiation.Furthermore,osteogenic differentiation were ohserved by using ALP and ARS staining.Results:Prostate cancer cell exosomes can enter pre-osteoblasts.Compared to control cells,the expression of Osx,Runx2 and Alp are decreased in pre-osteoblasts cells treated with prostate cancer cell exosomes(P<0.05).The staining results showed that osteogenic differentiation was inhibited.Conclusion:Prostate cancer cells secrete exosomes that can act on pro-osteoblasts and inhibit osteoblast differentiation.