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人脐带间充质干细胞抑制前列腺癌转移的作用及机理研究

The Inhibitory Effect and Mechanism of Human Umbilical Cord Mesenchymal Stem Cells on Prostate 

  • 摘要: 目的观察人脐带间充质干细胞(UMSCs)是否能抑制前列腺癌的生长和转移,并探讨其机制。方法 以组织贴壁法分离人UMSCs。UMSCs与LNCaP、PC-3前列腺癌细胞Transwell共培养12 h、24 h、48 h和72 h,检测前列腺癌细胞的增殖状态,计算共培养72 h时UMSCs对前列腺癌细胞增殖的抑制率。UMSCs与LNCaP、PC-3前列腺癌细胞Transwell共培养48 h,检测UMSCs对前列腺癌细胞侵袭力的抑制情况,计算抑制率。采用MILLIPLEX MILLIPLEX?MAP液相芯片技术检测共培养72 h后培养上清液中基质金属蛋白酶(MMP)-2、MMP-9和基质金属蛋白酶组织抑制因子(TIMP)-1、TIMP-2的表达。结果与UMSCs共培养后,前列腺癌细胞的增殖受到了抑制。LNCaP增殖速度在共培养72 h时低于对照组(P<0.05),细胞增殖抑制率为37.21%;PC-3增殖速度在共培养48 h和72 h时低于对照组(P<0.05),72 h时细胞增殖抑制率为31.27%。Transwell侵袭实验结果提示共培养48 h后,前列腺癌细胞侵袭力受到抑制,侵袭力抑制率分别为48.35%(LNCaP)和46.91%(PC-3)。共培养72 h后,LNCaP和PC-3分泌的MMP-2、MMP-9较对照组减少(P<0.05),TIMP-1、TIMP-2的表达较对照组增加(P<0.05)。结论UMSCs可通过分泌MMPs的拮抗剂TIMPs来抑制前列腺癌细胞的增殖和侵袭转移能力。

     

    Abstract: Objective To test whether umbilical cord mesenchymal stem cells (UMSCs) can inhibit the growth and metastasis of prostate cancer cells, and to investigate the mechanism. Methods The UMSCs from human umbilical cord tissue were isolate by explant technique. After being co-cultured the UMSCs with LNCaP and PC-3 cells for 24 h, 48 h and 72 h, LNCaP and PC-3 cells’ proliferation were tested and the 72 h proliferation inhibitory rate (IR) was calculated. Transwell invasion assay was used to test the migration and invasion abilities of prostate cancer cells after being co-cultured with UMSCs for 48 h. The invasion IR were calculated. The expression of matrix metalloproteinases (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 in the 72 h co-culture supernatants were tested by MILLIPLEX?MAPmethod. Results The proliferation of prostate cancer cells was inhibited after being co-cultured with UMSCs. The proliferation rate of LNCaP was lower than control group at 72 h (P<0.05), and the proliferation IR was 37.21%. The proliferation rate of PC-3 was lower than control group at 48 h and 72 h (P<0.05), and the proliferation IR was 31.27% at 72 h. Transwell invasion assays showed that co-culturing 48 h with UMSCs inhibited the invasive abilities of LNCaP and PC-3, and the invasion IR were 48.35% (LNCaP) and 46.91% (PC-3). Co-culturing 72 h, the expression of MMP-2 and MMP-9 were down-regulated (P<0.05) and the secretion of TIMP-1 and TIMP-2 were up-regulated (P<0.05) compared with control group. Conclusion UMSCs can inhibit the proliferation and invasion abilities of prostate cancer cells by secreting TIMPs, the antagonist of MMPs, which suppressed the overexpression of MMPs.

     

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