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  

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

作　　者：刘太阳 李杰 沈雁冰 闫泽晨[2] 桂琦 文秀华 侯铁奇 Liu Taiyang;Li Jie;Shen Yanbing;Yan Zechen;Gui Qi;Wen Xiuhua;Hou Tieqi(Department of Urology,Zhumadian Central Hospital,Zhumadian 463000,China;Department of Urology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]驻马店市中心医院泌尿外科,驻马店463000 [2]郑州大学第一附属医院泌尿外科,郑州450052

出　　处：《中华泌尿外科杂志》2022年第1期67-72,共6页Chinese Journal of Urology

摘　　要：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;蛋白质印迹法检测Caspase 3、脾酪氨酸激酶(Syk)、Ki-67表达。将Syk过表达载体质粒及其阴性对照分别转染至T24细胞中,转染后再分别用100 mg/L蒲葵子提取物处理,按上述方法检测细胞存活率、克隆形成数量及细胞凋亡率。用不同浓度蒲葵子提取物处理裸鼠膀胱癌模型,观察裸鼠一般情况和肿瘤生长情况;免疫组化染色检测膀胱癌组织Caspase 3、Syk、Ki-67蛋白表达。结果蒲葵子低、中、高剂量组较对照组T24细胞存活率显著降低[(88.50±3.65)%、(70.58±2.47)%、(48.90±2.37)%与(98.25±4.26)%],克隆形成数量显著减少[(101.33±3.40)、(84.00±2.94)、(60.00±2.16)个与(121.33±4.64)个],细胞凋亡率显著升高[(11.45±0.59)%、(17.71±0.64)%、(21.33±0.83)%与(7.86±0.43)%],Ki-67蛋白表达水平显著降低,Caspase3、Syk蛋白表达水平显著升高,且均呈浓度依赖性(均P<0.05)。pcDNA3.1-Syk组较pcDNA3.1组细胞存活率显著降低[(63.87±2.53)%与(98.45±3.54)%],克隆形成数量显著减少[(74.33±2.87)个与(121.33±3.68)个],细胞凋亡率显著升高[(18.39±0.63)%与(7.89±0.45)%](均P<0.05)。pcDNA3.1-Syk+蒲葵子高剂量组较pcDNA3.1+蒲葵子高剂量组的细胞存活率显著降低[(29.80±1.63)%与(49.33±2.76)%],克隆形成数量显著减少[(33.00±2.94)个与(59.67±3.30)个],细胞凋亡率显著升高[(26.93±0.68)%与(21.25±0.78)%](均P<0.05)。膀胱癌模型裸鼠实验结果显示,对照组和蒲葵子低、中、高剂量组裸鼠移植瘤体积分别为(1209.75±64.37)、(1006.31±40.49)、(530.58±42.87)、(267.58±16.73)mm^(3),移植瘤重量分别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

关 键 词：膀胱肿瘤 蒲葵子提取物 脾酪氨酸激酶 增殖 凋亡 

分 类 号：R285[医药卫生—中药学]