SERPINE1影响结肠癌细胞RKO和LoVo的增殖与凋亡  

Effects of SERPINE1 on Proliferation and Apoptosis of Colon Adenocarcinoma Cells RKO and LoVo

作　　者：陈德合 李登玉 郭刚 王杰 周倩 李洪涛 CHEN Dehe;LI Dengyu;GUO Gang;WANG Jie;ZHOU Qian;LI Hongtao(Department of General Surgery,the Fifth People's Hospital of Shangqiu City,Shangqiu Henan 476000,China)

机构地区：[1]商丘市第五人民医院普外科,河南商丘476000 [2]商丘市第五人民医院超声科,河南商丘476000 [3]联勤保障部队940医院普外科 [4]兰州大学第二临床医学院

出　　处：《联勤军事医学》2024年第4期277-284,共8页Military Medicine of Joint Logistics

基　　金：甘肃省科技计划项目(22JR5RA005);部队专项培育面上项目(2021yxky021)。

摘　　要：目的探究丝氨酸蛋白酶抑制剂1(serine proteinase inhibitor family E member 1,SERPINE1)在结肠癌(colon adenocarcinoma,COAD)中的表达、意义及对COAD细胞增殖与凋亡的影响,了解其在COAD进展中发挥的生物学功能。方法通过Gene Expression Profilling Interactive Analysis2.0(GEPIA2.0)数据库分析SERPINE1在COAD中的表达与分期及预后关系。收集作者医院5例COAD患者癌组织与对应的正常结肠组织标本,应用Western blot实验验证SERPINE1在COAD中的表达;接着构建SERPINE1过表达与干扰表达的慢病毒载体,进行细胞转染,通过实时荧光定量逆转录聚合酶链反应(real-time reverse transcription quantitative polymerase chain reaction,RT-qPCR)和Western blot对实验细胞筛选并验证过表达和干扰表达的效率;进而通过细胞计数试剂盒(cell counting kit 8,CCK-8)和Hoechst细胞核染色检测细胞增殖与凋亡;最后通过Western blot实验了解其对增殖标志物Ki-67(marker of proliferation Ki-67,MKI67)、Bcl-2关联X蛋白(Bcl-2 associated X protein,Bax)与B细胞淋巴瘤2基因(B cell lymphoma 2,Bcl-2)比值的影响。结果通过GEPIA2.0数据库查找并分析COAD中SERPINE1表达与临床分期和预后关系发现,SERPINE1在COAD中表达水平高于正常结肠组织,SERPINE1的表达水平与临床分期正相关,而与预后生存总生存期负相关(P均<0.05)。CCK-8实验结果表明,oe-RKO组和oe-LoVo组COAD细胞在48h时间点吸光度(optical density,OD)值大于oe-RKO-NC组和oe-LoVo-NC组,sh-RKO-822组和sh-LoVo-822组COAD细胞在48 h时间点OD值小于sh-RKO-NC和sh-LoVo-NC组(P均<0.05)。Hoechst凋亡细胞核染色结果表明,oe-RKO组和oe-LoVo组COAD细胞凋亡数量低于oe-RKO-NC组和oe-LoVo-NC组,sh-RKO-822组和sh-LoVo-822组COAD细胞凋亡数量明显高于sh-RKO-NC组和sh-LoVo-NC组(P均<0.05)。Western blot结果显示,oe-RKO组和oe-LoVo组COAD细胞MKI67蛋白表达明显高于oe-RKO-NC组和oe-LoVo-NC组,Bax/Bcl-2明显低于oe-RKO-NC组和oe-LoVo-NC组;sh-Objective To explore the expression and significance of serine proteinase inhibitor family E member 1(SERPINE1) in colon adenocarcinoma(COAD) and its effects on proliferation and apoptosis of COAD cells, and to explore its biological function in the progression of COAD.Methods The relationship between SERPINE1 expression and clinical stage and prognosis of COAD was analyzed by Gene Expression Profilling Interactive Analysis2.0(GEPIA2.0) database. Cancer tissues and corresponding normal colonic tissues were collected from 5 patients with COAD, the expression of SERPINE1 in COAD was verified by Western blot;then SERPINE1 overexpression and interference lentiviral vectors were constructed for cell transfection, real-time reverse transcription quantitative polymerase chain reaction(RT-qPCR) and Western blot were used to screen the experimental cells and verify the overexpression and interference expression efficiency;cell proliferation and apoptosis were detected by cell counting kit 8(CCK-8) and Hoechst nuclear staining;finally, the effects of the inhibitor on the marker of proliferation Ki-67(MKI67), Bcl-2 associated X protein(Bax) and B cell lymphoma 2(Bcl-2) gene ratio were detected by Western blot.Results The GEPIA2 database was used to search and analyze the relationship between the expression of SERPINE1 in COAD and clinical stage as well as prognosis, it was found that the expression level of SERPINE1 in COAD was higher than that in normal colonic tissues, the expression level of SERPINE1 was positively correlated with clinical stage, but negatively correlated with prognosis and overall survival(all P<0.05). The results of CCK-8 assay showed that the optical density(OD) value of COAD cells in oe-RKO group and oe-LoVo group at 48h was higher than that of oe-RKO-NC group and oe-LoVo-NC group, the OD value of COAD cells in sh-RKO-822 group and sh-LoVo-822 group at 48h was lower than that of sh-RKO-NC and sh-LoVo-NC groups(all P<0.05). Hoechst staining showed that the number of apoptotic COAD cells in oe-RKO grou

关 键 词：结肠癌 丝氨酸蛋白酶抑制剂1 增殖 凋亡 生物学功能 

分 类 号：R735.3[医药卫生—肿瘤]