lncRNA MSC-AS1通过调节miR-429/RhoA轴对人鼻咽癌细胞增殖、凋亡、迁移和侵袭的影响  

Impacts of LncRNA MSC-AS1 on the proliferation,apoptosis,migration and invasion of human nasopharyngeal carcinoma cells by regulating the miR-429/RhoA axis

作　　者：程莉雅[1] 吴洁 牛金明[1] CHENG Liya;WU Jie;NIU Jinming(Department of Otolaryngology,Weifang Hospital of Traditional Chinese Medicine,Weifang,Shandong 261000,China)

机构地区：[1]潍坊市中医院耳鼻喉科,山东潍坊261000

出　　处：《国际检验医学杂志》2023年第22期2789-2795,共7页International Journal of Laboratory Medicine

基　　金：潍坊市卫生健康委员会中医药科研项目[2022年(第4类)第072号]。

摘　　要：目的探索长链非编码RNA(lncRNA)MSC-AS1通过调节微小RNA(miR)-429/Ras同源基因家族成员A(RhoA)轴对人鼻咽癌(NPC)细胞增殖、凋亡、侵袭和迁移的影响。方法实时荧光定量PCR法(qPCR)检测NPC细胞系(CNE-1、HNE2、HONE-1细胞)以及人鼻咽上皮细胞系NP69细胞中lncRNA MSC-AS1、miR-429、RhoA mRNA的表达水平。以CNE-1细胞为研究对象,将si-MSC-AS1、miR-429 inhibitor、miR-429 mimics及相应阴性对照分别转染或共转染CNE-1细胞,记为对照组(未转染)、si-NC组、si-MSC-AS1组、mimics NC组、miR-429 mimics组、si-MSC-AS1+inhibitor NC组、si-MSC-AS1+miR-429 inhibitor组。MTT法、流式细胞术、Transwell分别检测细胞增殖、凋亡、迁移和侵袭能力;双荧光素酶实验分别验证miR-429与MSC-AS1、RhoA的靶向关系;免疫印迹法检测RhoA、B淋巴细胞因子相关x蛋白(Bax)、上皮细胞钙黏蛋白(E-cadherin)、细胞周期素D1(CyclinD1)、神经钙黏蛋白(N-cadherin)的表达水平。结果miR-429分别与MSC-AS1、RhoA存在靶向关系。与NP69细胞相比,CNE-1、HNE2、HONE-1细胞中MSC-AS1、RhoA mRNA的表达水平显著增加,miR-429表达水平显著降低,差异有统计学意义(P<0.05)。与对照组、si-NC组相比,si-MSC-AS1组细胞增殖率、迁移和侵袭数、MSC-AS1、N-cadherin、RhoA、CyclinD1比较表达水平显著下降,细胞凋亡率、E-cadherin、Bax显著增加,差异有统计学意义(P<0.05);与对照组、mimics NC组相比,miR-429 mimics组细胞增殖率、迁移和侵袭数、N-cadherin、RhoA、CyclinD1表达水平显著下降,细胞凋亡率、miR-429、E-cadherin、Bax显著增加,差异有统计学意义(P<0.05);抑制miR-429表达水平可减弱干扰MSC-AS1对细胞恶性生物学行为的影响(P<0.05)。结论干扰MSC-AS1可以抑制NPC细胞增殖、侵袭及迁移,促进其凋亡,可能与miR-429/RhoA轴有关。Objective To explore the impacts of long non-coding RNA(lncRNA)MSC-AS1 on the proliferation,apoptosis,invasion and migration of human nasopharyngeal carcinoma(NPC)cells by regulating the microRNA(miR)-429/Ras homologous gene family member A(RhoA)axis.Methods real time quantitative Pcr(qPCR)was performed to measure the expression levels of lncRNA MSC-AS1,miR-429 and RhoA mRNA in NPC cell lines(CNE-1,HNE2,HONE-1 cells)and human nasopharyngeal epithelial cell line NP69 cells.Taking CNE-1 cells as the research object,the CNE-1 cells were transfected or co-transfected with si-MSC-AS1,miR-429 inhibitor,miR-429 mimics and corresponding negative controls,which were recorded as control group(untransfected),si-NC group,si-MSC-AS1 group,mimics NC group,miR-429 mimics group,si-MSC-AS1+inhibitor NC group,and si-MSC-AS1+miR-429 inhibitor group.MTT method,flow cytometry and Transwell were performed to measure cell proliferation,apoptosis,migration and invasion abilities;dual luciferase experiments were performed to verify the targeting relationship of miR-429 with MSC-AS1 and RhoA;Western blot was performed to measure the expressions of RhoA,B-lymphokine-associated X protein(Bax),epithelial cell cadherin(E-cadherin),cyclin D1(CyclinD1)and neural cadherin(N-cadherin).Results MiR-429 had a targeting relationship with MSC-AS1 and RhoA,respectively.Compared with NP69 cells,the expressions of MSC-AS1 and RhoA mRNA in CNE-1,HNE2,and HONE-1 cells were significantly increased,and the expression of miR-429 was significantly decreased,and the differences were statistically significant(P<0.05).Compared with the control group and si-NC group,the cell proliferation rate,the numbers of migration and invasion,the expression level of MSC-AS1,N-cadherin,RhoA and CyclinD1 in si-MSC-AS1 group were significantly decreased,and the cell apoptosis rate,E-cadherin,and Bax were significantly increased,and the differences were statistically significant(P<0.05);compared with the control group and the mimics NC group,the cell proliferation rate,the number

关 键 词：人鼻咽癌细胞 增殖 长链非编码RNA MSC-AS1 凋亡 微小RNA-429/RhoA轴 

分 类 号：R739.63[医药卫生—肿瘤]