MPPa-PDT抑制骨肉瘤MG63细胞侵袭和迁移

发布时间：2018-05-08 05:14

本文选题：焦脱镁叶绿酸-a甲酯 + 光动力疗法　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:本研究以体外培养的骨肉瘤MG63细胞为研究对象,拟用MPPa介导的光动力疗法处理骨肉瘤MG63细胞,探讨焦脱镁叶绿酸-a甲酯介导的光动力疗法对人骨肉瘤MG-63细胞迁移及侵袭的影响及其可能机制。为MPPa-PDT用于骨肉瘤远处转移抑制及临床治疗提供实验理论基础。方法:将对数生长期骨肉瘤MG-63细胞分为空白对照组(Control组)、单纯MPPa药物组(MPPa组)、单纯光照组(LED组)以及MPPa-PDT实验组(MPPa-PDT组)。人骨肉瘤MG-63细胞经MPPa-PDT处理后采用CCK8检测不同时间点的细胞活性,明确MPPa-PDT对骨肉瘤MG63细胞的药物毒性效应;划痕实验及Transwell小室模型实验检测细胞迁移、侵袭能力,探明MPPa-PDT能否抑制骨肉瘤MG63细胞的迁移及侵袭能力;通过Western blot检测迁移侵袭标志性蛋白E-钙粘蛋白(E-cad)和基质金属蛋白酶(MMP)-2、-9蛋白的表达水平,以明确MPPa-PDT是否抑制了E-钙粘蛋白(E-cad)和基质金属蛋白酶(MMP)-2、-9蛋白的表达。结果:MPPa-PDT处理MG-63细胞以后,细胞活性明显降低,在12、24及48小时MPPa-PDT组细胞增殖能力较Control组、MPPa组、LED组存在明显抑制作用。通过MPPa-PDT处理后,我们可以观察到MPPa-PDT组细胞的划痕愈合能力、迁移活力及侵袭能力较Control组、MPPa组、LED组明显下降,MPPa-PDT对骨肉瘤MG-63细胞的迁移侵袭有抑制作用。Western blot检测结果则显示MPPa-PDT组的E-cad明显高于Control组、MPPa组及LED组,而MMP-2、MMP-9的表达则明显低于其他三组(P0.05),在蛋白水平,MPPa-PDT可对迁移侵袭关键蛋白起抑制作用。结论:MPPa-PDT抑制人骨肉瘤MG-63细胞的侵袭和迁移能力;上调E-cad的表达、下调MMP-2,-9的表达可能是其作用机制之一。
[Abstract]:Objective: in this study, osteosarcoma MG63 cells cultured in vitro were used to treat MG63 cells with MPPa mediated photodynamic therapy. To investigate the effect of methyl pyromagnesia-a photodynamic therapy on the migration and invasion of human osteosarcoma MG-63 cells and its possible mechanism. To provide experimental theoretical basis for the suppression of distant metastasis and clinical treatment of osteosarcoma by MPPa-PDT. Methods: MG-63 cells in logarithmic phase of osteosarcoma were divided into control group (control group), MPPa drug group (MPPA group), light group (LED group) and MPPa-PDT experimental group (MPPa-PDT group). Human osteosarcoma MG-63 cells treated with MPPa-PDT were treated with CCK8 to determine the cytotoxicity of MPPa-PDT to MG63 cells at different time points, scratch test and Transwell chamber model test were used to detect cell migration and invasion ability. To determine whether MPPa-PDT can inhibit the migration and invasion of osteosarcoma MG63 cells, and to detect the expression levels of E-cad- and MMP-2P protein by Western blot. In order to determine whether MPPa-PDT inhibits the expression of E-cadand and MMP-2P-9 protein of matrix metalloproteinases (MMP) and matrix metalloproteinase (MMP). Results the cell activity of MG-63 cells was significantly decreased after treated with 1: MPPa-PDT, and the proliferation ability of MPPa-PDT group was significantly inhibited than that of Control group at 24 and 48 hours. After treatment with MPPa-PDT, we can observe the ability of scratch healing of MPPa-PDT cells. Migration activity and invasion ability were significantly lower in Control group than those in Control group. Western blot analysis showed that E-cad in MPPa-PDT group was significantly higher than that in Control group and LED group. The expression of MMP-2 and MMP-9 was significantly lower than that of the other three groups, and MPPa-PDT could inhibit the key proteins of migration and invasion at the protein level. Conclusion the invasion and migration ability of human osteosarcoma MG-63 cells was inhibited by WMPPa-PDT, and the up-regulation of E-cad and the down-regulation of MMP-2 ~ (-9) expression may be one of the mechanisms of the inhibition of invasion and migration of human osteosarcoma MG-63 cells.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R738

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