香菇多糖对膀胱癌细胞增殖和迁移行为的影响

Effects of lentinan on the proliferation and migration of bladder cancer cells

目的观察香菇多糖(Lentinan)对膀胱癌UM-UC3细胞增殖和迁移行为的影响,初步探究其分子机制。方法采用不同浓度香菇多糖溶液培养UM-UC3细胞,通过噻唑蓝(MTT)实验、Transwell实验、划痕迁移实验和流式细胞术等分别评价细胞的增殖和迁移能力,通过Western blot实验检测相关标志物的蛋白表达量。结果香菇多糖可显著抑制膀胱癌UM-UC3细胞增殖和迁移,并呈现出一定的剂量和时间依赖性。与对照组(0μg/mL)比,低浓度组(20μg/mL)和高浓度组(60μg/mL)中G0/G1期细胞比例由(53.2±2.1)%分别上升至(57.2±0.9)%和(67.9±0.5)%,迁移细胞数由(308±30)个分别降低至(222±27)和(86±12)个,划痕愈合率分别由(56.2±2.4)%降低至(44.2±4.2)%和(19.4±2.9)%,差异均具有统计学意义(P<0.05)。免疫荧光染色结果显示:经香菇多糖处理后,周期蛋白Ki67荧光强度呈降低趋势。香菇多糖可抑制膀胱癌细胞上皮间充质转化基因(N-cadherin、Vimentin),周期相关基因(CDK2、CDK6)蛋白表达,上调原癌基因P53蛋白表达。结论香菇多糖可能通过周期阻滞和抑制上皮细胞间充质转化(EMT)途径抑制膀胱癌细胞增殖和迁移,在术前新辅助化疗中具有一定的应用潜力。

Objective To investigate the inhibitory effects of lentinan on the proliferation and migratio n of bladder cancer UM-UC3 cells and to explore the underlying mechanism.Methods UM-UC3 cells were co-incubated with lentinan at dif-ferent concentrations.The proliferation and migration were determined with MTT assay,Transwell assay,scratch assay and flow cytometry.The protein expressions of relevant markers were detected with Western blot.Results Lentinan inhibited the proliferation and migration of UM-UC3 cells in a dose-and time-dependent manner.Compared with blank control(0μg/mL),in the low concentration group(20μg/mL)and high concentration group(60μg/mL),the percentage of G0/G1 phase cells increased from(53.2±2.1)%to(57.2±0.9)%and(67.9±0.5)%,the number of migrated cells decreased from(308±30)to(222±27)and(86±12),and the healing rate of scratches decreased from(56.2±2.4)%to(44.2±4.2)%and(19.4±2.9)%,respectively(all P<0.05).The results of immunofluorescence indicated that Ki67 fluorescence intensity decreased obviously after lentinan treatment.Lentinan inhibited the expressions of N-cadherin,Vimentin,CDK2 and CDK6,but upregulated the expression of P53.Conclusion Lentinan can inhibit the proliferation and migration of bladder cancer cells by inhibiting cell cycle and epithelial-mesenchymal transition(EMT),which exhibits remarkable application potential in preoperative neoadjuvant chemotherapy.