松弛素通过 AKT/VEGF 通路促进人 Saos-2 骨肉瘤细胞的体外肿瘤生长、侵袭和血管生成。

Relaxin promotes in vitro tumour growth, invasion and angiogenesis of human Saos-2 osteosarcoma cells by AKT/VEGF pathway.

机构信息

Department of Spine, the Affiliated Hospital of Medical College, Qingdao University, China.

出版信息

Eur Rev Med Pharmacol Sci. 2013 May;17(10):1345-50.

Abstract

OBJECTIVES

In the present study, we determine the role of relaxin on cellular growth, invasion and angiogenesis of osteosarcoma Saos-2 cells in vitro, and discuss the molecular mechanisms of this action.

MATERIALS AND METHODS

Saos-2 cells were transfected with Akt1/2 siRNA or VEGF siRNA for 24 hours then treated with 10-100 ng/mL recombinant human relaxin-2 (rh-RLN) for 48 h. MTT, matrigel and bone marrow-derived endothelial cells (BMDECs) was used for cell proliferation, invasion and angiogenesis assay. Western blot was used for relaxin-2, pAKT and VEGF protein assay.

RESULTS

The results showed treatment with 10-100 ng/mL rh-RLN resulted in 18%, 48%, 107%, 212% increase in cell proliferation, respectively (vs control, *p < 0.05;**p < 0.01), the relative invasive cells was 1.4;1.9;2.6;4.8 (control was defined to 1) (vs control, #p < 0.01; ##p < 0.001) and the relative anglogenic branch points in Saos-2 cells was 1.04;1.36;1.69;2.10 (control was defined to 1.00) (vs control, *p < 0.05; **p < 0.01). Furthermore, treatment with rh-RLN exhibited a significant increase in the expression level of pAKT and VEGF proterin in dose-dependent manner. Saos-2 cells were transfected with AKT1/2 siRNA for 24 h. No significant increase of VEGF protein expression was shown after rh-RLN treatment.

CONCLUSIONS

These results suggested that rh-RLN could promoted proliferation, invasion and angiogenesis by upregulation pAKT-dependent VEGF expression.

摘要

目的

本研究旨在探讨松弛素（RLN）对体外人骨肉瘤 Saos-2 细胞的增殖、侵袭和血管生成的作用及其分子机制。

材料和方法

Saos-2 细胞转染 Akt1/2siRNA 或 VEGFsiRNA24 h 后，用 10-100 ng/ml 重组人松弛素-2（rh-RLN）处理 48 h。用 MTT、基质胶和骨髓来源的内皮细胞（BMDECs）进行细胞增殖、侵袭和血管生成实验。Western blot 检测 RLN、pAKT 和 VEGF 蛋白的表达。

结果

结果显示，10-100 ng/mlrh-RLN 处理组细胞增殖分别增加了 18%、48%、107%和 212%（与对照组相比，*p<0.05；**p<0.01），相对侵袭细胞数分别为 1.4、1.9、2.6 和 4.8（对照组定义为 1）（与对照组相比，#p<0.01；##p<0.001），Saos-2 细胞的相对血管生成分支数分别为 1.04、1.36、1.69 和 2.10（对照组定义为 1.00）（与对照组相比，*p<0.05；**p<0.01）。此外，rh-RLN 处理呈剂量依赖性增加 pAKT 和 VEGF 蛋白的表达水平。Saos-2 细胞转染 Akt1/2siRNA24 h 后，rh-RLN 处理后 VEGF 蛋白表达无明显增加。