Xu H, Shi H L, Hao J W, Shu K P, Zhang Y T, Hou T Q
Department of Urology, Henan People's Hospital, Zhengzhou 450000, China.
Medical College of Henan University, Kaifeng 475004, China.
Zhonghua Zhong Liu Za Zhi. 2022 Apr 23;44(4):334-340. doi: 10.3760/cma.j.cn112152-20200507-00420.
To explore the effect and mechanism of Casticin (CAS) on the proliferation, migration and invasion of bladder cancer T24 cells. T24 cells were cultured in vitro and divided into control group, 5, 10, 20 μmol/L CAS groups, si-NC group, si-TM7SF4 group, CAS+ pcDNA group and CAS+ pcDNA-TM7SF4 group. Cell counting kit-8 (CCK-8) was used to detect cell proliferation; Transwell was used to detect cell migration and invasion; western blot was used to detect the protein expressions of cyclin D1, p21, MMP-2, MMP-9 and TM7SF4, and real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to detect the expression of TM7SF4 mRNA. The inhibition rates of T24 cells in the 5, 10, 20 μmol/L CAS groups were (17.68±1.41)%, (33.54±3.16)% and (61.44±5.50)%, respectively, higher than (0.00±0.00)% of the control group (<0.001), but the numbers of migration and invasion were 72.83±5.66, 59.13±4.27, 41.25±3.22 and 55.83±5.15, 42.19±3.06, 31.13±3.22, respectively, lower than 86.11±5.16 and 68.82±5.29 of the control group (<0.001). The protein expression levels of cyclin D1, MMP-2, MMP-9, TM7SF4 and the expression levels of TM7SF4 mRNA in the 5, 10, and 20 μmol/L CAS groups were lower than the control group (<0.001). However, the protein expression levels of p21 were 0.37±0.03, 0.51±0.04, and 0.66±0.06, respectively, higher than 0.25±0.03 in the control group (<0.001). The inhibition rate of T24 cells in the si-TM7SF4 group was (50.35±4.67)%, higher than (6.31±0.58)% in the si-NC group (<0.001), but the numbers of migration and invasion were 53.51±4.18 and 42.92±3.81, lower than 85.26±4.99 and 67.93±4.64 of the si-NC group (<0.001). The protein expression levels of TM7SF4, CyclinD1, MMP-2, MMP-9 in the si-TM7SF4 group were lower than the si-NC group (<0.001). However, the protein expression level of p21 in the si-TM7SF4 group was higher than the si-NC group (<0.001). The inhibitory rate of T24 cells in the CAS+ pcDNA-TM7SF4 group was (21.45±2.46)%, lower than (64.06±4.49)% of the CAS+ pcDNA group (<0.001), but the number of migration and invasion in the CAS+ pcDNA-TM7SF4 group were 75.66±6.57 and 59.35±5.40, higher than 40.43±3.85 and 30.25±3.32 in the CAS+ pcDNA group (<0.001). The protein expression levels of TM7SF4, CyclinD1, MMP-2 and MMP-9 in the CAS+ pcDNA-TM7SF4 group were higher than the CAS+ pcDNA group (<0.001), but the protein expression level of p21 was lower than the CAS+ pcDNA group (<0.001). CAS may suppress the proliferation, migration and invasion of bladder cancer T24 cells by inhibiting the expression of TM7SF4.
探讨紫花牡荆素（CAS）对膀胱癌T24细胞增殖、迁移和侵袭的影响及其机制。体外培养T24细胞，分为对照组、5、10、20 μmol/L CAS组、si-NC组、si-TM7SF4组、CAS+ pcDNA组和CAS+ pcDNA-TM7SF4组。采用细胞计数试剂盒-8（CCK-8）检测细胞增殖；采用Transwell检测细胞迁移和侵袭；采用蛋白质免疫印迹法检测细胞周期蛋白D1、p21、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）和TM7SF4的蛋白表达，采用实时定量逆转录聚合酶链反应（RT-qPCR）检测TM7SF4 mRNA的表达。5、10、二十 μmol/L CAS组T24细胞的抑制率分别为（17.68±1.41）%、（33.54±3.16）%和（61.44±5.50）%，高于对照组的（0.00±0.00）%（<0.001），但迁移和侵袭细胞数分别为72.83±5.66、59.13±4.27、41.25±3.——22和55.83±5.15、42.19±3.06、31.13±3.22，低于对照组的86 .11±5.16和68.82±5.29（<0.001）。5、10、二十 μmol/L CAS组细胞周期蛋白D1、MMP-2、MMP-9、TM7SF4的蛋白表达水平及TM7SF4 mRNA的表达水平均低于对照组（<0.001）。然而，p21的蛋白表达水平分别为0.37±0.03、0.51±0.04和0.66±0.06，高于对照组的0.25±0.03（<0.001）。si-TM7SF4组T24细胞的抑制率为（50.35±4.67）%，高于si-NC组的（6.31±0.58）%（<0.001），但迁移和侵袭细胞数分别为53.51±4.18和42.92±3.81，低于si-NC组的85.26±4.99和67.93±4.64（<0.001）。si-TM7SF4组TM7SF4、细胞周期蛋白D1、MMP-2、MMP-9的蛋白表达水平低于si-NC组（<0.001）。然而，si-TM7SF4组p21的蛋白表达水平高于si-NC组（<0.001）。CAS+ pcDNA-TM7SF4组T24细胞的抑制率为（21.45±2.46）%，低于CAS+ pcDNA组的（64.06±4.49）%（<0.001），但CAS+ pcDNA-TM7SF4组的迁移和侵袭细胞数分别为75.66±6.57和59.35±5.40，高于CAS+ pcDNA组的40.43±3.85和30.25±3.32（<0.001）。CAS+ pcDNA-TM7SF4组TM7SF4、细胞周期蛋白D1、MMP-2和MMP-9的蛋白表达水平高于CAS+ pcDNA组（<0.001），但p21的蛋白表达水平低于CAS+ pcDNA组（<0.001）。CAS可能通过抑制TM7SF4表达来抑制膀胱癌T24细胞的增殖、迁移和侵袭。
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