摘要
目的探讨长链非编码RNA(long non-coding RNA,lncRNA)双特异性磷酸酶5假基因1(dual-specificity phosphatase 5 pseudogene 1,DUSP5P1)对三阴性乳腺癌(triple negative breast cancer,TNBC)细胞生物学行为的作用。方法采用实时荧光定量PCR法检测TNBC细胞系MDA-MB-231、4T1、MDA-MB-468及正常人乳腺导管上皮细胞MCF-10A中lncRNA DUSP5P1的表达水平;将MDA-MB-231细胞系分为对照组、sh-vector组和sh-DUSP5P1组,分别不感染慢病毒、只感染空载质粒及感染sh-DUSP5P1-慢病毒(lentivirus,lenti)。采用MTT实验和克隆形成实验检测细胞增殖能力,Transwell实验检测细胞侵袭和迁移能力,Western Blot实验检测上皮标志物E-cadherin和间质标志物Vimentin、Snail的表达。结果TNBC细胞系MDA-MB-231、4T1及MDA-MB-468中lncRNA DUSP5P1的表达水平均明显高于正常人乳腺导管上皮细胞系MCF-10A(F=65.10,P<0.05)。细胞培养12、24、48、72 h后,sh-DUSP5P1组吸光度值(OD 490值)明显低于对照组和sh-vector组(分别F=33.62,90.72,41.17,77.94;均P<0.05)。细胞培养7 d后,与对照组和sh-vector组比较,sh-DUSP5P1组克隆形成数明显下降(F=575.1,P<0.05)。敲减DUSP5P1后,TNBC细胞系MDA-MB-231侵袭和迁移能力明显下降(分别F=933.30,160.20;均P<0.05),上皮标志物E-cadherin明显上升(F=6.21,P<0.05),间质标志物Vimentin和Snail明显下降(分别F=100.11,227.37;均P<0.05)。结论LncRNA DUSP5P1可能通过上皮间质转化过程促进TNBC细胞侵袭和迁移。
Objective To investigate the effect of long non-coding RNA(lncRNA)dual-specificity phosphatase 5 pseudogene 1(DUSP5P1)on the biological behavior of triple negative breast cancer(TNBC)cells.Methods The expression of lncRNA DUSP5P1 in the TNBC cell lines MDA-MB-231,4T1,MDA-MB-468 and normal human mammary epithelium cell line MCF-10A were dectected by quantitative real time polymerase chain reaction(qRT-PCR).The MDA-MB-231 cells were divided into negative control(NC)group,negative shRNA-Lenti(vector group)or Lenti-sh-DUSP5P1(sh-DUSP5P1 group).The cell proliferation ability was determined by MTT assay and cloning formation experiment,migration and invasion ability were measured by Transwell assay,the epithelial marker E-cadherin and interstitial marker Vimentin,Snail were dectected by Western Blot.Results The relative expression levels of lncRNA DUSP5P1 in TNBC cell lines MDA-MB-231,4T1and MDA-MB-468 were significantly higher than those in normal human mammary epithelium cell line MCF-10A(F=65.10,P<0.05).MTT assay and cloning formation experiment demonstrated that the proliferation ability of sh-DUSP5P1 group was lower than NC group and sh-vector group(F=33.62,90.72,41.17,77.94 and 575.1;respectively P<0.05).Transwell assay showed that the invasion and migration ability of sh-DUSP5P1 group was lower than NC group and sh-vector group(F=933.30 and 160.20;respectively P<0.05).After DUSP5P1 was knocked down,the epithelial marker E-cadherin in MDA-MB-231 was increased significantly(F=6.21,P<0.05),but the interstitial marker Vimentin and Snail decreased significantly(F=100.11 and 227.37;respectively P<0.05).Conclusion LncRNA DUSP5P1 may promote invasion and migration of TNBC cells through epithelial-mesenchymal transition process.
作者
曹亚伟
桂百卉
CAO Yawei;GUI Baihui(Department of Laboratory,the Huaian First People’s HospitalAffiliated to Nanjing Medical University,Huaian Jiangsu 223001,China)
出处
《转化医学杂志》
2020年第1期12-16,共5页
Translational Medicine Journal
关键词
三阴性乳腺癌
长链非编码RNA
双特异性磷酸酶5假基因1
Triple negative breast cancer(TNBC)
Long non-coding RNA(lncRNA)
Dual-specificity phosphatase 5 pseudogene 1(DUSP5P1)
