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目的探讨西红花酸对人成骨肉瘤MG63细胞增殖、凋亡和迁移能力的影响及其作用机制。方法以0、50、100、200、400μmol/L不同浓度西红花酸处理人成骨肉瘤MG63细胞48 h后,用倒置显微镜观察MG63细胞形态的变化;用MTT法检测其对MG63存活率的影响;用划痕实验观察其对MG63细胞迁移能力的影响;用Western blot法检测其对MG63细胞中bcl-2、Caspase-3、P53、P21等凋亡相关蛋白表达水平的影响。结果不同浓度西红花酸作用48h后,倒置显微镜观察到MG63细胞呈现凋亡的形态结构;MTT结果显示随着西红花酸浓度从0增加到400μmol/L,MG63细胞的存活率由(100±9.5)%减少为(26.8±4)%,且与阴性对照组比较差异有统计学意义(P<0.01);划痕实验可见经西红花酸处理细胞后MG63细胞的迁移能力降低;Western blot结果提示不同浓度经西红花酸(0、50、100、200μmol/L)作用MG63细胞后,其bcl-2/β-actin的灰度比值从(56.09±4.36)%降至(18.78±1.20)%,p21/β-actin、Caspase-3/β-actin、P53/β-actin的灰度比值分别由(28.61±1.71)%、(13.90±1.65)%、(21.18±1.95)%增至(71.37±2.46)%、(112.36±2.97)%、(97.43±3.48)%,与对照组相比差异均有统计学意义(P<0.01),且均呈浓度依赖性。结论西红花酸能够促进MG63细胞凋亡并抑制其增殖,其机制可能与抑制凋亡基因bcl-2表达下调和促进凋亡的基因caspase3、p53、p21基因表达上调的调节有关。
Objective To investigate the effect of crocetin on the proliferation, apoptosis and migration of human osteosarcoma MG63 cells and its mechanism. Methods Human osteosarcoma MG63 cells were treated with 0, 50, 100, 200 and 400μmol / L crocetin for 48 h. The morphology of MG63 cells was observed by inverted microscope. The survival rate of MG63 cells was determined by MTT assay Effects of different concentrations on the migration of MG63 cells were evaluated by scratch test. The expression of bcl-2, Caspase-3, P53 and P21 in MG63 cells was detected by Western blot. Results After 48 h of crocetin exposure, the morphology of apoptotic MG63 cells was observed by inverted microscope. The MTT results showed that the survival rate of MG63 cells increased from (0) to 400 μmol / L ± 9.5%) to (26.8 ± 4)%, and there was a significant difference compared with the negative control group (P <0.01). Scratch experiment showed that the migration ability of MG63 cells decreased after treatment with crocetin; The result of blot showed that the gray value of bcl-2 / β-actin decreased from (56.09 ± 4.36)% to (18.78 ±) in MG63 cells treated with different concentrations of crocetin (0,50,100,200μmol / The gray ratios of p21 / β-actin, Caspase-3 / β-actin and P53 / β-actin increased from 28.61 ± 1.71%, 13.90 ± 1.65% and 21.18 ± 1.95% (71.37 ± 2.46)%, (112.36 ± 2.97)% and (97.43 ± 3.48)%, respectively, which were significantly different from the control group (P <0.01). Conclusion Crocetin can promote the apoptosis of MG63 cells and inhibit the proliferation of MG63 cells. The mechanism may be related to the up-regulation of the expression of caspase3, p53 and p21 genes, which may down-regulate the expression of apoptosis-related genes bcl-2 and promote apoptosis.