RPRD1B对三阴性乳腺癌细胞生物学功能及上皮间质转化的影响  

Effect of RPRD1B on biological function and epithelial mesenchymal transition of triple-negative breast cancer

作　　者：李维妙 郑国旭 梁亮[1] 马宇光[1] 魏星 LI Weimiao;ZHENG Guoxu;LIANG Liang;MA Yuguang;WEI Xing(Department of Oncology,Second Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710004,China;Department of Immunology,Air force Medical University;Department of Gynecology and Obstetrics,Second Affiliated Hospital of Xi′an Jiaotong University)

机构地区：[1]西安交通大学第二附属医院肿瘤科,西安710004 [2]空军军医大学基础医学院免疫学教研室 [3]西安交通大学第二附属医院妇产科

出　　处：《山西医科大学学报》2024年第5期544-552,共9页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(82203675);陕西省重点研发计划项目(2020SF-115)。

摘　　要：目的探讨核前mRNA结构域调节因子1B(RPRD1B)对三阴性乳腺癌(TNBC)细胞增殖、凋亡、侵袭、迁移及上皮间质转化(EMT)的影响。方法免疫组化检测RPRD1B在三阴性乳腺癌组织和癌旁组织的表达水平,Western blot检测三阴性乳腺癌细胞系MDA-MB-231和HCC-1937中RPRD1B的表达水平。三阴性乳腺癌细胞系MDA-MB-231和HCC-1937分别用sh-RPRD1B慢病毒(sh-RPRD1B组)和sh-NC慢病毒(sh-NC组)感染,然后检测下调效率;CCK-8法检测细胞增殖能力,流式细胞术检测细胞凋亡水平,Transwell实验检测细胞侵袭能力,划痕实验检测细胞迁移能力,Western blot和qRT-PCR法检测敲低RPRD1B对三阴性乳腺癌细胞中上皮间质转化标志性蛋白E-cadherin和N-cadherin的影响。结果RPRD1B在三阴性乳腺癌组织和三阴性乳腺癌MDA-MB-231细胞系和HCC-1937细胞系中均表达。CCK-8结果显示,与sh-NC组比较,sh-RPRD1B组MDA-MB-231和HCC-1937细胞增殖能力显著降低(P<0.01)。流式细胞术结果显示,与sh-NC组比较,sh-RPRD1B组细胞凋亡率显著上升(P<0.01)。Transwell结果显示,与sh-NC组比较,sh-RPRD1B组细胞侵袭能力显著降低(P<0.01)。划痕实验结果显示,与sh-NC组比较,sh-RPRD1B组细胞迁移能力显著降低(P<0.01)。与sh-NC组比较,sh-RPRD1B组细胞上皮间质转化标志分子E-cadherin表达水平升高,N-cadherin表达水平降低。结论敲除RPRD1B表达使三阴性乳腺癌细胞的凋亡能力增强而增殖能力减弱,其侵袭和迁移能力的减弱可能与上皮间质转化过程相关。Objective To investigate the effects of regulatory nuclear pre-mRNA domain containing 1B(RPRD1B)on the proliferation,apoptosis,invasion,migration and epithelial-mesenchymal transition(EMT)of triple-negative breast cancer(TNBC).Methods The expression level of RPRD1B in TNBC tissues and adjacent non-tumor tissues was detected by immunohistochemistry.The expression level of RPRD1B in TNBC cell lines MDA-MB-231 and HCC-1937 was detected by Western blot.The TNBC cell lines MDA-MB-231 and HCC-1937 were respectively transfected with sh-RPRD1B lentivirus(sh-RPRD1B group)and sh-NC lentivirus(sh-NC group).The downregulation efficiency was detected.CCK-8 assay was used to detect the cell proliferation.Flow cytometry was used to detect the cell apoptosis.Transwell assay was used to detect the cell invasion.The wound healing assay was used to detect the cell migration.Western blot and qRT-PCR were used to detect the expressions of EMT-related proteins(E-cadherin and N-cadherin)in TNBC cells.Results RPRD1B was expressed in TNBC tissues and in human TNBC cell lines,MDA-MB-231 and HCC-1937.CCK-8 analysis showed that the proliferation ability in sh-RPRD1B group was significantly decreased compared with sh-NC group(P<0.01).Flow cytometry analysis showed that the apoptosis rate in sh-RPRD1B group was significantly increased compared with sh-NC group(P<0.01).Transwell analysis showed that the invasion ability in sh-RPRD1B group was significantly reduced compared with sh-NC group(P<0.01).The wound healing analysis showed that the migration ability in sh-RPRD1B group was significantly decreased compared with sh-NC group(P<0.01).The expression level of EMT marker E-cadherin was elevated while the expression of N-cadherin was reduced in sh-RPRD1B group compared with sh-NC group.Conclusion Knockdown of RPRD1B could increase the apoptosis of TNBC cells and reduce the proliferation,which may be related to the epithelial mesenchymal transformation process.

关 键 词：三阴性乳腺癌 核前mRNA结构域调节因子 细胞增殖 细胞侵袭 上皮间质转化 

分 类 号：R737.9[医药卫生—肿瘤]