山慈菇多糖对膀胱癌细胞迁移、侵袭及miR-411表达的影响

Effects of polysaccharides extracted from rhizoma pleionis on migration,invasion and expression of miR-411 of bladder cancer cells

目的考察山慈菇多糖对膀胱癌细胞的迁移和侵袭及miR-411表达的影响。方法膀胱癌细胞T24分为对照组(正常培养)、低、中、高剂量实验组(1,2,4 mg·mL^(-1)山慈菇多糖处理)、miR-411组(转染miR-411 mimics)和miR-NC组(转染miR-NC)。用Lipofectamine;2000转染试剂,在T24细胞中转染miR-411 mimics和其阴性对照miR-NC。以碘化丙啶单染法评估各组膀胱癌T24细胞的周期情况;以Transwell小室法测定各组膀胱癌T24细胞的迁移和侵袭情况;以蛋白质印迹法分析各组膀胱癌T24细胞的P21蛋白、细胞周期蛋白D1、细胞核相关抗原Ki-67、E钙黏蛋白、N钙黏蛋白的表达情况;以实时定量反转录聚合酶链反应测定各组膀胱癌T24细胞的miR-411基因表达水平。结果对照组、低、中、高剂量实验组、miR-NC组和miR-411组T24细胞G0-G1细胞比例分别为32.95±0.76,37.00±0.83,43.03±0.86,48.07±0.83,32.96±0.73和40.32±0.80;S期细胞比例分别为33.60±0.74,29.45±0.82,23.30±0.81,18.84±0.78,33.60±0.74和25.81±0.84;迁移细胞数分别为173.67±1.70,152.00±1.63,122.33±1.70,84.33±1.25,174.67±1.70和138.00±1.63;侵袭细胞数分别为97.00±1.63,79.67±1.25,65.67±0.94,45.00±0.82,95.67±2.05和73.00±1.63;miR-411基因表达水平分别为0.99±0.07,1.34±0.10,1.72±0.11,1.97±0.13,1.00±0.03和1.56±0.12。低、中、高剂量实验组与对照组比较,差异均有统计学意义(均P<0.05);miR-411组与miR-NC组比较,差异均有统计学意义(均P<0.05)。结论山慈菇多糖抑制膀胱癌细胞的迁移侵袭,可能与其上调miR-411的表达有关。

ObjectiveTo investigate the effects of polysaccharide extracted from the Rhizoma Pleionis on the proliferation,migration and invasion of bladder cancer cells and the expression of miR-411.Methods Bladder cancer cells T24 were divided into control group,experimental-L,experimental-M and experimental-H group (1,2,4 mg·mL^(-1) of polysaccharide extracted from the Rhizoma Pleionis),miR-411 group (transfected with miR-411 mimics) and miR-NC group (transfected miR-NC).Using Lipofectamine;2000 transfection reagent,miR-411 mimics and its negative control miR-NC were transfected in T24 cells.The propidium iodide single staining method was used to evaluate the cycle of bladder cancer T24 cells;the Transwell chamber method was used to measure the migration and invasion of bladder cancer T24 cells in each group;Western blot was used to detect P21,Cyclin D1,nuclear associated antigen Ki67,E-cadherin,N-cadherin,protein expression;expression of miR-411 gene in bladder cancer T24 cells in each group was measured by qRT-PCR.Results The proportions of G0-G1 of T24 cells in the control group,experimental-L group,experimental-M group,experimental-H group,miR-NC group and miR-411 group were 32.95±0.76,37.00±0.83,43.03±0.86,48.07±0.83,32.96±0.73 and 40.32±0.80,respectively;the proportion of S-phase cells were33.60±0.74,29.45±0.82,23.30±0.81,18.84±0.78,33.60±0.74 and 25.81±0.84;the number of migrating cells was 173.67±1.70,152.00±1.63,122.33±1.70,84.33±1.25,174.67±1.70 and138.00±1.63;the number of invasive cells was 97.00±1.63,79.67±1.25,65.67±0.94,45.00±0.82,95.67±2.05 and 73.00±1.63,respectively;the expression levels of miR-411 gene were 0.99±0.07,1.34±0.10,1.72±0.11,1.97±0.13,1.00±0.03 and 1.56±0.12.Compared between the control group and the experimental-L,-M,-H groups,the differences were statistically significant (all P<0.05);Compared between the miR-411 group and the miR-NC group,the differences were statistically significant (all P<0.05).Conclusion Polysaccharide extracted from the Rhizoma Pleionis could inhibit the proliferation,migration and invasion of bladder cancer cells,which may be related to up-regulate the expression of miR-411.