膜联蛋白A1调控膀胱癌细胞增殖、凋亡及迁移的机制  

The mechanism of Annexin A1 regulating the proliferation,apoptosis and migration of bladder cancer cells

作　　者：王涛[1] 白培德 谢顺强 孙安冉 邢金春[1] 陈斌[1] Wang Tao;Bai Peide;Xie Shunqiang;Sun Anran;Xing Jinchun;Chen Bin(Department of Urology,The First Affiliated Hospital of Xiamen University,Xiamen 361001,China)

机构地区：[1]厦门大学附属第一医院泌尿外科,361001

出　　处：《中华泌尿外科杂志》2019年第12期932-936,共5页Chinese Journal of Urology

基　　金：福建省自然科学基金项目(2017J01355);厦门市科技计划项目(3502Z 20184014)。

摘　　要：目的探讨膜联蛋白A1(ANXA1)在膀胱癌细胞增殖、凋亡和迁移中的作用及调控机制。方法2018年2月至2019年6月,以膀胱癌T24细胞为模型,取对数生长期的T24细胞进行转染,根据转染的质粒将细胞分为过表达对照组(ctrl组,转染pLV-Ctrl)、ANXA1过表达组(ANXA1组,转染ANXA1-OE)、敲降对照组(shctrl组,转染pLL3.7-shCtrl)、ANXA1敲降组1(shANXA1-1组,转染ANXA1-sh1)、ANXA1敲降组2(shANXA1-2组,转染ANXA1-sh2)和ANXA1敲降组3(shANXA1-3组,转染ANXA1-sh3),转染24 h后收集细胞用于实验。采用荧光定量PCR法检测ANXA1的mRNA表达水平;采用CCK-8法检测细胞的活性;采用流式细胞术检测细胞凋亡和周期;采用Transwell法检测细胞的迁移情况。结果荧光定量PCR检测结果显示,ANXA1组和ctrl组的ANXA1表达量分别为15369.00±874.20和1.00±0.07,差异有统计学意义(P<0.001);shANXA1-2组、shANXA1-3组、shctrl组的ANXA1表达量分别为0.51±0.04、0.51±0.02、1.00±0.04,差异有统计学意义(P<0.001)。ANXA1组和ctrl组的细胞活性分别为2.04±0.02和1.61±0.01,差异有统计学意义(P<0.001);shANXA1-2组、shANXA1-3组、shctrl组的细胞活性分别为1.40±0.002、1.31±0.003、1.73±0.01,差异有统计学意义(P<0.001)。流式细胞检测结果显示,ANXA1组和ctrl组G0/G1期细胞数量分别为28.14±0.33和46.19±0.73,差异有统计学意义(P<0.001),shANXA1-2组、shANXA1-3组、shctrl组的G0/G1期细胞数量分别为58.67±0.49、62.34±4.01、45.59±0.19,shANXA1-2组、shANXA1-3组与shctrl组比较差异均有统计学意义(P<0.001,P<0.05)。ANXA1组和ctrl组细胞凋亡率差异无统计学意义(P>0.05),而shANXA1-2组、shANXA1-3组、shctrl组的细胞凋亡率分别为13.04%、14.58%、7.76%,差异有统计学意义(P<0.001)。细胞功能实验结果显示,ANXA1组和ctrl组迁移细胞数量分别为(525.00±9.30)个和(385.70±13.40)个,差异有统计学意义(P<0.01),shANXA1-2组、shANXA1-3组、shctrl组迁移细胞数量分别为(214.70±6.40)、(2Objective Explore the function and regulatory mechanism of Annexin A1(ANXA1)in bladder cancer cell proliferation,apoptosis and migration.Methods From February 2018 to June 2019,we use T24 cells as the model and divide it into over-expression control group(ctrl),ANXA1 over-expression group(ANXA1),knockdown control group(shctrl),ANXA1 knockdown group 1(shANXA1-1),ANXA1 knockdown group 2(shANXA1-2)and ANXA1 knockdown group 3(shANXA1-3).24 hours after the culture,the cells were collected and the mRNA expression level of ANXA1 was detected by Real-Time quantitative PCR.The cell activity was detected by CCK-8;the cell apoptosis and cycle were detected by flow cytometry.The cell migration was detected by Transwell assay.Results The Real-Time quantitative PCR showed that the expression of ANXA1 in the over expression group was significantly higher than that in the over expression control group(15369.00±874.20 and 1.00±0.07,P<0.001).The expression of ANXA1 in the knockdown group 2 and 3 were significantly lower than that in the knockdown control group(0.51±0.04,0.51±0.02 and 1.00±0.04,P<0.001).Compared with the over expression control group(1.61±0.01),the cell activity of the over expression group(2.04±0.02)was significantly increased(P<0.001),while the activity of the knockdown group 2 and 3(1.40±0.002 and 1.31±0.003)were significantly decreased than the knockdown ctrl group(1.73±0.01)(P<0.001).The results of flow cytometry showed that the number of G0/G1 cells in the over-expression group was significantly lower than that in the over-expression control group(28.14±0.33 and 46.19±0.73,P<0.001),while that in the knockdown group 2 and 3 were significantly higher than that in the knockdown control group(58.670±0.49,62.34±4.01 and 45.59±0.19,P<0.001 and P<0.05).There was no significant difference in the number of apoptosis between the over-expression group and the over-expression control group(P>0.05),while the number of apoptosis in the knockdown group 2 and 3 were significantly higher than that in the knockd

关 键 词：膜联蛋白A1 T24细胞 ANXA1 荧光定量PCR法 流式细胞检测 荧光定量PCR检测 膀胱癌细胞 细胞活性 

分 类 号：R73[医药卫生—肿瘤]