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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12h、24h、48h和72h,检测前列腺癌细胞的增殖状态,计算共培养72h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48h,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○RMAP液相芯片技术检测共培养72h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72h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48h和72h时低于对照组(P<0.05),72h时细胞增殖抑制率为31.27%。Transwell侵袭实验结果提示共培养48h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72h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。
Objective To observe whether human umbilical cord mesenchymal stem cells (UMSCs) can inhibit the growth and metastasis of prostate cancer and to explore its mechanism. Methods Tissue adherent method was used to separate human UMSCs. UMSCs were co-cultured with LNCaP and PC-3 prostate cancer cells Transwell for 12h, 24h, 48h and 72h respectively to detect the proliferation of prostate cancer cells. The inhibition rate of UMSCs on the proliferation of prostate cancer cells was calculated 72h after co-culture. UMSCs were cocultured with LNCaP and PC-3 prostate cancer cells Transwell for 48 hours to detect the inhibitory effect of UMSCs on invasiveness of prostate cancer cells. The inhibition rate was calculated. The expression of matrix metalloproteinase (MMP) -2, MMP-9 and TIMP-1, TIMP-2 in culture supernatants were detected by MILLIPLEX ¡Á RMAP liquid-phase microarray. Results After co-cultured with UMSCs, the proliferation of prostate cancer cells was inhibited. The proliferation rate of LNCaP was lower than that of the control group at 72h (P <0.05), and the cell proliferation inhibition rate was 37.21%. The proliferation rate of PC-3 was lower than that of the control group at 48h and 72h (P <0.05) Cell proliferation inhibition rate was 31.27%. The results of Transwell invasion assay indicated that the invasiveness of prostate cancer cells was inhibited 48 hours after co-culture and the inhibitory rates of invasiveness were 48.35% (LNCaP) and 46.91% (PC-3), respectively. After co-cultured for 72h, the expressions of MMP-2 and MMP-9 in LNCaP and PC-3 were significantly decreased (P <0.05) and the expressions of TIMP-1 and TIMP-2 in control group were higher than those in control group (P <0.05). Conclusion UMSCs can inhibit the proliferation, invasion and metastasis of prostate cancer cells by secreting TIMPs, an antagonist of MMPs.