干扰LncRNA BCYRN1对乳腺癌MCF-7细胞增殖、运动和移植瘤生长的影响  

Effect of LncRNA BCYRN1 on Proliferation and Mobility of MCF-7 Breast Cancer Cells and Xenograft Growth in Mice

作　　者：吉棚 赵华山 王晨 刘淑荣 马建增 JI Peng;ZHAO Huashan;WANG Chen;LIU Shurong;MA Jianzeng(Department of Breast and Thyroid Surgery,Sunshine Union Hospital,Weifang,Shandong,261000,China;Department of Oncology,Sunshine Union Hospital,Weifang,Shandong,261000,China)

机构地区：[1]阳光融和医院乳腺甲状腺外科,山东省潍坊市261000 [2]阳光融和医院肿瘤内科,山东省潍坊市261000

出　　处：《医学分子生物学杂志》2023年第5期444-449,共6页Journal of Medical Molecular Biology

基　　金：潍坊市卫生健康委员会科研项目(No.WFWSJK-2021-314)。

摘　　要：目的探讨短发卡RNA(short hairpin RNA,shRNA)干扰长链非编码RNA BCYRN1(long noncoding RNA BCYRN1,LncRNA BCYRN1)对乳腺癌MCF-7细胞增殖、运动及移植瘤生长的影响。方法实时定量PCR(real-time quantitative PCR,RT-PCR)检测乳腺癌及癌旁组织BCYRN1表达,将乳腺癌MCF-7细胞分为空白对照组(Control)、阴性对照组(shRNA-NC)和干扰组(sh-BCYRN1),通过试剂盒转染转入目的片段并进行RT-PCR验证,Edu法、transwell法、划痕实验和蛋白质印迹法(Western blotting)分别检测细胞增殖、侵袭、迁移能力及其相关蛋白表达,免疫荧光检测细胞波形蛋白(Vimentin)水平。通过裸鼠腋下注射MCF-7细胞建立裸鼠移植瘤模型,待成瘤后将裸鼠分为空白对照组、干扰组,每组各20只,干扰组、空白对照组裸鼠分别给予sh-BCYRN1及其阴性对照转染干预,观察4周后移植瘤BCYRN1表达、肿瘤重量和Ki67、N-钙粘蛋白(N-cadherin)蛋白表达。结果乳腺癌组织BCYRN1表达高于癌旁组织(P<0.05)。比较空白对照组,干扰组Edu阳性细胞百分比、单个视野细胞侵袭数目、细胞迁移率、Vimentin阳性率降低(P<0.05),Ki67、N-cadherin蛋白表达降低,E-钙粘蛋白(E-cadherin)蛋白表达升高(P<0.05)。干扰组移植瘤BCYRN1表达降低,肿瘤质量、Ki67及N-cadherin蛋白表达低于空白对照组移植瘤(P<0.05)。结论shRNA干扰LncRNA BCYRN1可抑制乳腺癌MCF-7细胞增殖、运动和移植瘤生长。Objective To explore the effect of LncRNA BCYRN1 on the proliferation and motility of MCF-7 breast cancer cells and the growth of xenograft in mice by using short hairpin RNA(shRNA)interfering.Methods The expression level of BCYRN1 in breast cancer and para-cancerous tissues was detected by real-time quantitative PCR(RT-PCR).MCF-7 cells were divided into 3 groups,the blank control group(Control),the negative control group(shRNA-NC)and the interference group(sh-BCYRN1).The target fragments were transfected,their effect on expressions of the target genes were then verified by RT-PCR.The cell proliferation,invasion,migration ability and the expression levels of related proteins were detected by Edu,transwell assay,wound-healing assay and Western blotting,respectively.Vimentin level was detected by immunofluorescence method.The nude mice were given subcutaneous injection of MCF-7 cells to construct xenograft models.After tumor formation,nude mice were divided into two groups,the blank control group(negative control)and the interference group(sh-BCYRN1),and there were 20 cases in each group.The tumor weight and the expression levels of BCYRN1,Ki67 and N-cadherin in the transplanted tumors were detected 4 weeks later.Results The expression level of BCYRN1 was higher in breast cancer tissues than in para-cancerous tissues(P<0.05).The percentage of Edu positive cells,the number of migrated and invasion cells,the positive rate of Vimentin,and the expression levels of Ki67 and N-cadherin were decreased in the interference group when compared with the blank control group(P<0.05),while the expression level of E-cadherin was increased(P<0.05).In the interference group,the expression level of BCYRN1 was decreased,and the tumor mass,the expression levels of Ki67 and N-cadherin were lower than those in the blank control group(P<0.05).Conclusion Interfering of LncRNA BCYRN1 can inhibit the proliferation and motility of MCF-7 breast cancer cells and the growth of xenograft.

关 键 词：长链非编码RNA BCYRN1 MCF-7乳腺癌细胞 细胞增殖 移植瘤 

分 类 号：R737.9[医药卫生—肿瘤]