抑制lncRNA HIF1A-AS1表达对长春新碱耐药视网膜母细胞瘤细胞株裸鼠移植瘤生长及转移的影响  

Influences of inhibiting the expression of lncRNA HIF1A⁃AS1 on the transplanted tumor growth and metastasis of vincristine⁃resistant retinoblastoma cell line in nude mice

作　　者：何道侗 高玉 HE Daotong;GAO Yu(Department of Ophthalmology,First Affiliated Hospital of Naval Medical University(Changhai Hospital),Shanghai 200433,China)

机构地区：[1]海军军医大学第一附属医院(长海医院)眼科,上海200433

出　　处：《临床肿瘤学杂志》2023年第9期785-791,共7页Chinese Clinical Oncology

基　　金：上海市自然科学基金项目(19ZR1456300)。

摘　　要：目的探讨抑制长链非编码RNA(lncRNA)缺氧诱导因子1α-反义RNA1(HIF1A-AS1)表达对长春新碱(VCR)耐药视网膜母细胞瘤(Rb)细胞株裸鼠的生存期、移植瘤生长和转移的影响。方法体外培养人Rb细胞株HXO-RB44、Y79、SO-RB50,通过逐步增加VCR浓度的方法刺激诱导建立其相应的VCR耐药Rb细胞HXO-RB44/VCR、Y79/VCR、SO-RB50/VCR,以实时荧光定量PCR(qPCR)实验检测上述细胞中lncRNA HIF1A-AS1、缺氧诱导因子1α(HIF-1α)表达。于裸鼠背部皮下注射Y79/VCR细胞悬液建立VCR耐药Rb细胞株裸鼠模型,随机分为模型组、VCR组、VCR+lncRNA HIF1A-AS1敲低组、VCR+lncRNA HIF1A-AS1空载组,以VCR和相应质粒分组干预处理后,检测各组裸鼠移植瘤的肿瘤体积、抑瘤率、转移情况及存活率;qPCR实验检测各组裸鼠移植瘤lncRNA HIF1A-AS1及HIF-1α、多药耐药1(MDR1)、乳腺癌相关耐药蛋白(BCRP/ABCG2)mRNA表达;免疫印迹法检测VCR耐药Rb细胞株裸鼠移植瘤HIF-1α、P-gp、ABCG2蛋白表达。结果与人Rb正常细胞HXO-RB44、Y79和SO-RB50相比,lncRNA HIF1A-AS1、HIF-1α在其相应VCR耐药细胞HXO-RB44/VCR、Y79/VCR、SO-RB50/VCR中的表达均显著升高(P<005)。与模型组比较,VCR+lncRNA HIF1A-AS1敲低组VCR耐药Rb细胞株裸鼠移植瘤肿瘤体积、转移器官数、转移灶总数、lncRNA HIF1A-AS1及HIF-1α、MDR1、ABCG2 mRNA表达、HIF-1α及P-gp、ABCG2蛋白表达均显著降低(P<005),裸鼠给药后30 d存活率、抑瘤率显著升高(P<005);VCR组、VCR+lncRNA HIF1A-AS1空载组VCR耐药Rb细胞株裸鼠各指标无显著变化(P>005)。与VCR组比较,VCR+lncRNA HIF1A-AS1敲低组VCR耐药Rb细胞株裸鼠移植瘤肿瘤体积、转移器官数、转移灶总数、lncRNA HIF1A-AS1及HIF-1α、MDR1、ABCG2 mRNA表达、HIF-1α及P-gp、ABCG2蛋白表达均显著降低(P<005),裸鼠给药后30 d存活率、抑瘤率显著升高(P<005);VCR+lncRNA HIF1A-AS1空载组VCR耐药Rb细胞株裸鼠各指标无显著变化(P>005)。结论下调lncRNA HIF1A-AS1表Objective To investigate the impacts of inhibiting the expression of long non-coding RNA(lncRNA)hypoxia-inducible factor 1α-antisense RNA1(HIF1A-AS1)on survival,transplanted tumor growth and metastasis of vincristine(VCR)-resistant retinoblastoma(Rb)cell line in nude mice.Methods Human Rb cell lines HXO-RB44,Y79,SO-RB50 were cultured in vitro,the corresponding VCR-resistant Rb cells HXO-RB44/VCR,Y79/VCR,SO-RB50/VCR were stimulated and established by gradually increasing the concentration of VCR,the expressions of lncRNA HIF1A-AS1 and hypoxia-inducible factor 1α(HIF-1α)in the above cells were detected by real-time fluorescence quantitative(qPCR)experiment.The VCR-resistant Rb cell line in nude mouse model was established by subcutaneous injection of Y79/VCR cell suspension on the back of nude mice,and they were randomly grouped into model group,VCR group,VCR+lncRNA HIF1A-AS1 knockdown group,and VCR+lncRNA HIF1A-AS1 empty load group.After grouping and intervention with VCR and corresponding plasmids,the tumor volume,tumor inhibition rate,metastasis and survival rate of transplanted tumors in nude mice in each group were measured.The mRNA expressions of lncRNA HIF1A-AS1 and HIF-1α,multidrug resistance 1(MDR1)and breast cancer-related drug resistance protein(BCRP/ABCG2)were detected by qPCR in nude mice transplanted tumors in each group.The protein expressions of HIF-1α,P-gp and ABCG2 in the transplanted tumor of VCR-resistant Rb cell line in nude mice were measured by western blotting.Results Compared with human Rb normal cells HXO-RB44,Y79 and SO-RB50,the expressions of lncRNA HIF1A-AS1 and HIF-1αwere significantly increased in their corresponding VCR-resistant cells HXO-RB44/VCR,Y79/VCR and SO-RB50/VCR(P<005).Compared with the model group,the transplanted tumor volume,number of metastatic organs,total number of metastatic foci,the mRNA expressions of lncRNA HIF1A-AS1,HIF-1α,MDR1 and ABCG2,and the protein expressions of HIF-1α,P-gp and ABCG2 in transplanted tumor of VCR-resistant Rb cell line in nude mice i

关 键 词：视网膜母细胞瘤 缺氧诱导因子1α-反义RNA1 耐药 生存期 移植瘤 生长转移 

分 类 号：R739[医药卫生—肿瘤]