Hsa-miR-145联合槲皮素对人前列腺癌LNCaP细胞侵袭、迁移的影响

Effect of Hsa-miR-145 combined with quercetin on invasion and migration of human prostate cancer cell line LNCaP

目的:研究Hsa-miR-145(人微小RNA-145,miR-145)联合槲皮素对人前列腺癌(LNCaP)细胞侵袭、迁移的影响,并探讨其机制。方法:将一定浓度的miR-145模拟物经脂质体包裹转染前列腺癌(LNCaP)细胞,并联合不同浓度的槲皮素,采用MTT法检测单用miR-145、单用槲皮素及miR-145联合槲皮素对前列腺癌细胞增殖的抑制作用,流式细胞术检测细胞早期凋亡率,Transwell及伤口愈合实验检测细胞侵袭、迁移的能力,Western blot检测细胞蛋白表达水平。结果:单用槲皮素的IC50为104.5μmol/L,槲皮素与随机序列联合使用IC50为87.69μmol/L,槲皮素与miR-145模拟物联合使用IC50为37.92μmol/L,药物增敏倍数为2.75。miR-145联合槲皮素促进LNCaP细胞早期凋亡,其早期凋亡率达11.4%,miR-145模拟物联合槲皮素作用于LNCaP细胞24、48小时后,伤口愈合率分别为16.7%、34.8%。槲皮素联合miR-145模拟物组侵袭细胞数为(26±3)个,而转移数为(56±4)个。结论:Hsa-miR-145能增强LNCaP细胞对槲皮素的敏感性,槲皮素组联合miR-145模拟物能促进LNCaP细胞早期凋亡,并能显著抑制LNCaP细胞侵袭、迁移的能力,其机制可能与下调E-cadherin、snail蛋白,上调N-cadherin蛋白的表达有关。

Objective： To study the effect of Hsa-miR-145（ human mircroRNA-145,miR-145） combined with quercetin on invasion and migration of human prostate cancer（ LNCa P） cells and to explore its potential molecular mechanism. Methods： A certain concentration of miR-145 was transfected into prostate cancer（ LNCa P） cells by liposome,and combined with different concentrations of quercetin. The effect of quercetin or Hsa-miR-145 alone and the combination of the two on the proliferation of LNCa P cells was detected with MTT. The effect on cell cycle was detected by flow cytometry. The cell invasion and migration were detected by Transwell and wound healing assay. The capability of cell colony formation was detected by a colony formation experiment. The level of cell protein expression was detected by Western blot. Results： IC50 of quercetin was 104. 5 μmol/L. The IC50 was 87. 69 μmol/L when combined use of quercetin and random sequence. The IC50 was 37. 92 μmol/L when combined use of quercetin and miR-145 mimics,the increased susceptibility multiples was 2. 75. miR-145 combined with quercetin promotes the early apoptosis of LNCap cells,and the early apoptosis rate of cells were 11. 4%. The wound healing rates were 16. 7% and34. 8% after miR-145 mimics combined with quercetin for 24 h,48 h. Quercetin combined with miR-145 in the number of invasive cells was 56 ± 4. Conclusion： Hsa-miR-145 could enhance the sensitivity of LNCa P cells to quercetin,quercetin group analogue of miR-145 could promote the early apoptosis of LNCa P cells. It could significantly inhibit LNCa P cell invasion and migration ability,the mechanism may be down-regulation of E-cadherin,snail protein,up-regulated expression of N-cadherin protein.