蒲葵子提取物对膀胱癌细胞增殖凋亡的影响及其相关机制

目的:探讨蒲葵子提取物对膀胱癌细胞增殖、凋亡的影响及其相关机制。方法:将T24细胞分别用含蒲葵子提取物且终浓度分别为0、25、50、100 mg/L的培养液培养，分别记为对照组和蒲葵子低、中、高剂量组。用细胞计数试剂盒8(CCK-8)检测细胞存活率；克隆形成实验检测细胞克隆形成数量；流式细胞术检测细胞凋亡；蛋白质印迹法...

Effect of extract of livistona chinensis on the proliferation and apoptosis of bladder cancer cells and its related mechanisms

Objective To study the effect of extract of livistona chinensis on the proliferation and apoptosis of bladder cancer cells and the related mechanism.Methods T24 cells were cultured in medium with the final concentration of 0,25,50 and 100 mg/L livistona chinensis extract,respectively.And then they were divided into control group and low,medium and high dose groups.The cell survival rate was detected by cell counting kit 8(CCK-8).Colony formation assay was used to detect the number of cell clones.Apoptosis was detected by flow cytometry.Western blot was used to detect the expression of related proteins.The Syk overexpression vector plasmid and its negative control were transfected into T24 cells.After transfection,the cells were treated with 100 mg/L livistona chinensis.The cell survival rate,colony formation number and apoptosis rate were detected by the above method.The bladder cancer model nude mice were treated with different concentrations of livistona chinensis extract.Under the microscope,the expression of protein was detected by immunohistochemical staining of bladder tissue.Results Compared with the control group,the survival rate of T24 cells in the low,medium and high dose livistona chinensis extract groups were significantly decreased(88.50±3.65)%,(70.58±2.47)%,(48.90±2.37)%vs.(98.25±4.26)%],and the number of clone formation decreased significantly(101.33±3.40),(84.00±2.94),(60.00±2.16)vs.(121.33±4.64)],and the apoptosis rate was significantly increased(11.45±0.59)%,(17.71±0.64)%,(21.33±0.83)%vs.(7.86±0.43)%].The expression level of Ki-67 protein was significantly decreased,while the expression levels of Caspase3 and Syk protein were significantly increased in a concentration dependent manner(P<0.05).The cell survival rate of pcDNA3.1-Syk group was significantly lower than that of pcDNA3.1 group(63.87±2.53)%vs.(98.45±3.54)%],the number of clone formation decreased significantly(74.33±2.87)vs.(121.33±3.68)],and the apoptosis rate was significantly increased(18.39±0.63)%vs.(7.89±0.45)%](all P<0.05).The cell survival rate in the high-dose group of livistona chinensis+pcDNA3.1-Syk was significantly lower than that in the high-dose group of livistona chinenisi+pcDNA3.1 group(29.80±1.63)%vs.(49.33±2.76)%],the number of clone formation decreased significantly(33.00±2.94)vs.(59.67±3.30)],and the apoptosis rate was significantly increased(26.93±0.68)%vs.(21.25±0.78)%](P<0.05).The experimental results of nude mice of bladder cancer model showed that the tumor volume of transplanted bladder cancer nude mice in the control group and the low,medium,and high dose livistona chinensis extract groups were(1209.75±64.37),(1006.31±40.49),(530.58±42.87),(267.58±16.73)mm³,respectively,the weight of the transplanted tumor were(0.36±0.08),(0.30±0.04),(0.26±0.03),(0.18±0.06)g,and the differences between the two groups were statistically significant(P<0.05).Immunohistochemical staining results showed that the expression of Sky and Caspase3 was increased and the expression of Ki-67 was decreased in the middle and high dose groups compared with that in the control group.Conclusion Extract of livistona chinensis can inhibit the proliferation and promote apoptosis of bladder cancer cells by up regulating Syk expression.