脂肪酸合成酶抑制剂TVB-2640对膀胱癌细胞的影响研究

Effects of fatty acid synthase inhibitor TVB-2640 in bladder cancer cell lines

目的研究脂肪酸合成酶(FASN)抑制剂TVB-2640对膀胱癌UMUC3细胞株增殖、凋亡、侵袭及转移的影响。方法取对数生长期UMUC3细胞,分为对照组(加入PBS)和低、中、高3个浓度实验组(12. 50,25,50μmol·L-1TVB-2640),分别处理24,48,72 h后用于后续实验。用噻唑蓝(MTT)法、克隆形成实验检测细胞增殖情况,以流式细胞术检测细胞凋亡情况,以细胞划痕实验检测细胞迁移能力变化,以Transwell实验检测细胞侵袭能力变化。结果药物处理24h后,低、中、高3个浓度实验组细胞增殖抑制率分别为(21. 97±1. 39)%,(28. 12±2. 22)%,(41. 67±2. 12)%,药物处理48 h后,细胞增殖抑制率分别为(30. 83±1. 92)%,(38. 12±1. 44)%,(62. 33±2. 32)%,药物处理72 h后,细胞增殖抑制率分别为(43. 22±2. 54)%,(58. 33±3. 52)%,(83. 32±3. 32)%,中、高浓度组与低浓度组比较,差异均有统计学意义(均P <0. 05)。TVB-2640作用于UMUC3细胞在24,48,72 h的IC50值分别为58. 92,40. 52,19. 41μmol·L-1,说明抑制效应呈时间依赖性。药物处理24 h后,25,50,100μmol·L-1实验组细胞凋亡率分别为(16. 72±0. 74)%,(21. 83±1. 11)%,(27. 40±1. 22)%,药物处理48 h后,细胞凋亡率分别为(23. 61±1. 12)%,(28. 33±1. 77)%,(30. 11±2. 24)%,各实验组细胞与对照组相比,差异均有统计学意义(均P <0. 05),高、低浓度实验组比较,差异均有统计学意义(均P <0. 05)。中、高浓度实验组处理细胞24 h,细胞平均迁移能力分别为16. 82±1. 57,79. 04±2. 11,各实验组细胞与对照组相比,差异均有统计学意义(均P <0. 05),说明药物对细胞的迁移活性具有显著抑制作用;中、高浓度实验组处理细胞24 h,细胞穿膜细胞数分别为117. 25±2. 11,66. 70±2. 01,各实验组细胞的侵袭能力与对照组(175. 10±2. 14)相比,差异均有统计学意义(均P <0. 05),说明药物对细胞的侵袭活性具有显著抑制作用。结论 FASN抑制剂TVB-2640可显著抑制膀胱癌UMUC3细胞的增殖、侵袭转移活性并诱导其凋亡,FASN可能是潜在的膀胱癌治疗靶点,FASN抑制剂有望成为一种新的膀胱癌治疗方法。

Objective To investigate the effects of fatty acid synthase(FASN)inhibitor TVB-2640 on proliferation,apoptosis,invasion and migration of bladder cancer UMUC3 cell lines.Methods The logarithmic growth phase UMUC3 cells were divided into control group(add PBS solution)and low,medium,high concentration experimental groups(TVB-2640,12.50,25,50μmol·L-1)and treated for 24,48,72 h.Cell proliferation was detected by MTT assay and colony formation assay.Cell apoptosis was determined by flow cytometry.Cell migration and invasion were detected by cell scratch assay and Transwell assay respectively.Results After administration TVB-2640 for 24 h,the inhibition rate of cell proliferation in the low,medium,high concentration experimental groups were(21.97±1.39)%,(28.12±2.22)%,(41.67±2.12)%;after treatment for 48 h,the inhibition rate of cell proliferation were(30.83±1.92)%,(38.12±1.44)%,(62.33±2.32)%;after treatment for 72 h,the inhibition rate of cell proliferation were(43.22±2.54)%,(58.33±3.52)%,(83.32±3.32)%.The IC50 values of the TVB-2640 on UMUC3 cells at 24,48,72 h were 58.92,40.52,19.41μmol·L-1,respectively,the TVB-2640 dramatically decreases UMUC3 proliferation in a dose-dependent and time-dependent manner.After treatment for 24 h with TVB-2640,the apoptosis rate in 25,50,100μmol·L-1 concentration experimental groups were(16.72±0.74)%,(21.83±1.11)%,(27.40±1.22)%;and after treatment for 48 h,the apoptosis rates were(23.61±1.12)%,(28.33±1.77)%,(30.11±2.24)%;the difference was significant compared with control group(P<0.05).After treatment for 24 h with TVB-2640,the average migration ability of medium and high concentration experimental groups were16.82±1.57,79.04±2.11,the TVB-2640 had a significant inhibitory effect on the cell migration activity(P<0.05).After treatment for 24 h,the transmembrane cells of medium and high concentration experimental groups were 117.25±2.11 and 66.70±2.01,indicating the TVB-2640 had a significant inhibitory effect on the cell invasion activity,compared with control group,which was 175.10±2.14(P<0.05).Conclusion FASN inhibitor TVB-2640 can significantly inhibit the proliferation,invasion and migration activity of bladder cancer UMUC3 cells and induce its apoptosis.FASN may be a potential therapeutic target for bladder cancer,FASN inhibitor is expected to be a new treatment for bladder cancer.