NRSN2-AS1在卵巢癌组织中表达的临床意义及体外生物学效应

Clinical significance of NRSN2-AS1 expression in ovarian cancer and its biological effect in vitro

目的探究长链非编码RNA(long non-coding RNA,lncRNA)NRSN2-AS1在卵巢癌组织中表达的临床意义及体外生物学效应。方法临床收集84例卵巢癌组织及相应正常癌旁组织,实时荧光定量PCR(RT-qPCR)技术检测NRSN2-AS1表达,并分析其与患者临床资料的关系;体外培养人卵巢上皮细胞HOSEpiC、人卵巢癌细胞A2780及OVCAR3,于A2780及OVCAR3细胞中分别转染过表达及干扰NRSN2-AS1载体及相应的对照载体,作为本研究的空白组、过表达组及对照组、干扰组;CCK-8实验检测细胞增殖能力的变化;Transwell侵袭及迁移实验检测细胞转移能力的变化,RT-qPCR技术及蛋白质印记(Western blot)技术检测细胞NRSN2-AS1潜在下游基因SRY相关HMG盒转录因子12(SOX12)mRNA及蛋白的表达。结果NRSN2-AS1在卵巢癌组织及细胞中显著高表达(P<0.05),且其高表达与患者预后不良、淋巴结转移及高临床分期显著相关(P<0.05);空白组及过表达组NRSN2-AS1表达分别为:1.00±0.08及5.78±0.41,SOX12 mRNA表达分别为:1.00±0.10及3.08±0.23;SOX12蛋白表达分别为:1.00±0.08及7.26±0.39,每视野侵袭细胞数分别为:22.7±4.9个及79.0±6.2个,每视野迁移细胞数为26.5±4.1个及43.5±4.5个;对照组及干扰组NRSN2-AS1表达分别为:1.00±0.11及0.37±0.04,SOX12 mRNA表达分别为:1.00±0.07及0.59±0.05;SOX12蛋白表达分别为:1.00±0.07及0.36±0.03,每视野侵袭细胞数68.3±6.1及30.6±5.5,每视野迁移细胞数为85.2±7.0个及22.7±4.2。相比空白组,过表达组SOX12 mRNA及蛋白表达均显著增高(P<0.05),细胞增殖及转移能力显著增强(P<0.05);相比对照组,干扰组SOX12 mRNA及蛋白表达均显著降低(P<0.05),细胞增殖及转移能力显著减弱(P<0.05)。结论NRSN2-AS1在卵巢癌组织中表达上调,且与患者预后不良及病情进展密切相关,体外可促进SOX12的表达及细胞的恶性行为。

Objective To investigate the clinical significance and biological effect of long non-coding RNA(lncRNA)NRSN2-AS1 in ovarian cancer.Methods The expression of NRSN2-AS1 was detected by real-time quantitative PCR(RT-qPCR)in 84 cases of ovarian cancer tissues and corresponding normal adjacent tissues,and the relationship between NRSN2-AS1 expression and clinical data of patients was analyzed.Human ovarian epithelial cells HOSEpiC and human ovarian cancer cells A2780 and OVCAR3 were cultured in vitro,and overexpression or interference of NRSN2-AS1 and control plasmids were respectively transfected into A2780 and OVCAR3 cells as the blank group,overexpression group and control group,interference group.CCK-8 assay was used to detect cell proliferation,while transwell invasion and migration assay were used to detect the change of cell metastasis ability,RT-qPCR and Western blot were used to detect the expressions of SRY related HMG box transcription factor 12(SOX12),a potential downstream gene of NRSN2-AS1.Results The expressions of NRSN2-AS1 in ovarian cancer tissues and cells were significantly higher than normal adjacent tissues,and the high expression of NRSN2-AS1 was significantly correlated with poor prognosis,lymph node metastasis and high clinical stage(P<0.05).In the blank and overexpression group,the expressions of NRSN2-AS1 were 1.00±0.08 and 5.78±0.41,the expressions of SOX12 mRNA were 1.00±0.10 and 3.08±0.23,the expression of SOX12 protein were 1.00±0.08 and 7.26±0.39,the invasion cells per field were 22.7±4.9 and 79.0±6.2,and the migration cells per field were 26.5±4.1 and 43.5±4.5.In the control and interference group,the expression of NRSN2-AS1 were 1.00±0.11 and 0.37±0.04,the expressions of SOX12 mRNA were 1.00±0.07 and 0.59±0.05,the expression of SOX12 protein were 1.00±0.07 and 0.36±0.03,the invasion cells per field were 68.3±6.1 and 30.6±5.5,and the migration cells per field were 85.2±7.0 and 22.7±4.2.Compared with the blank group,the expression of SOX12 mRNA and protein in the overexpression group was significantly increased(P<0.05),and the cell proliferation and metastasis ability were significantly enhanced(P<0.05).Compared with the control group,the expression of SOX12 mRNA and protein in the interference group was significantly decreased,and the ability of cell proliferation and metastasis was suppressed significantly(P<0.05).Conclusion The expression of NRSN2-AS1 is up-regulated in ovarian cancer,which is closely related to poor prognosis and progression of ovarian cancer.NRSN2-AS1 can promote the expression of SOX12 and the malignant behavior of ovarian cancer cells in vitro.