长链非编码RNAARHGAP5-AS1抑制乳腺癌细胞的增殖、迁移和侵袭

Long non-coding RNA ARHGAP5-AS1 inhibits the proliferation, migration and invasion of breast cancer cells

目的:检测长链非编码RNA(long non-coding RNA,lncRNA)ARHGAP5-AS1在乳腺癌组织及细胞中的表达,分析其表达与患者临床病理参数及预后的相关性,并初步探讨其对乳腺癌细胞体外增殖、迁移和侵袭的影响。方法:通过对TCGA数据库中乳腺癌相关数据集的生物信息学分析,筛选出在乳腺癌中低表达且与患者不良预后相关的lncRNA ARHGAP5-AS1,采用qPCR方法在江南大学附属医院肿瘤科从2010年4月至2016年10月收集的乳腺癌组织中验证其表达。采用χ2检验分析ARHGAP5-AS1表达与乳腺癌患者临床病理参数之间的关系,Kaplan-Meier生存分析构建生存曲线,比较高、低表达组的总生存期和无复发生存期。CCK-8实验、划痕实验和Transwell实验分别检测ARHGAP5-AS1敲低对乳腺癌细胞MDA-MB-231和BT-549的增殖、迁移和侵袭的影响。结果:TCGA数据库分析结果显示,ARHGAP5-AS1在乳腺癌组织中的表达水平显著低于正常乳腺组织(P<0.01),其低表达与较大肿瘤直径(T3)、远处转移(M1)、ER和PR阴性以及较短的总生存期显著相关(均P<0.05)。乳腺癌组织中ARHGAP5-AS1表达水平显著低于癌旁组织(P<0.05),其低表达与较大的肿瘤直径和淋巴结转移相关(均P<0.05)。同样,ARHGAP5-AS1在6株人乳腺癌细胞系(MDA-MB-231、BT-549、MDA-MB-468、MCF-7、HCC1937、Hs578T)中的表达水平也显著低于正常乳腺上皮细胞系(MCF-10A)(均P<0.05)。细胞功能实验显示,ARHGAP5-AS1敲低促进MDA-MB-231和BT-549细胞的增殖、迁移和侵袭(均P<0.05)。结论:ARHGAP5-AS1异常低表达可能通过促进乳腺癌细胞的增殖、迁移和侵袭影响乳腺癌的发生发展。

Objective: To detect the expression of long non-coding RNA(lncRNA) ARHGAP5-AS1 in breast cancer(BC) tissues and cell lines, analyze its relationship with the clinicopathological parameters and prognosis of BC patients, and to investigate its effect on the proliferation, migration and invasion of BC cells in vitro. Methods: LncRNA ARHGAP5-AS1 that low expressed in BC and associated with poor prognosis was screened out by bioinformatics analysis of BC-related data sets in TCGA database. qPCR was used to verify the expression of lncRNA ARHGAP5-AS1 in BC tissues collected from April 2010 to October 2016 in the Department of Oncology, Affiliated Hospital of Jiangnan University. χ2 test was used to analyze the relationship between the expression of ARHGAP5-AS1 and the clinicopathological parameters of BC patients. Kaplan-Meier survival analysis was performed to construct a survival curve to compare the overall and recurrence-free survival of the high and low expression groups. The effects of ARHGAP5-AS1 knockdown on the proliferation, migration and invasion of MDA-MB-231 and BT-549 cells were examined by CCK-8, Wound-healing assay and Transwell assay. Results: TCGA database analysis showed that the expression level of ARHGAP5-AS1 in BC tissues was significantly lower than that in normal breast tissues(P<0.01), and its low expression was significantly related to larger tumor diameter(T3), distant metastasis(M1), negative ER and PR, and shorter overall survival(all P<0.05). ARHGAP5-AS1 expression was significantly down-regulated in BC tissues compared with adjacent specimens(P<0.05), which was significantly associated with larger tumor diameter and lymph node metastasis(all P<0.05) in BC patients. Furthermore, ARHGAP5-AS1 was also reduced in 6 BC cell lines(MDA-MB-231, BT-549, MDA-MB-468, MCF-7, HCC1937, Hs578 T) compared with normal breast epithelial cell line MCF-10 A(all P<0.05). Cell function experiments showed that ARHGAP5-AS1 knockdown promoted proliferation, migration and invasion of MDA-MB-231 and BT-549 cells(all P<0.05). Conclusion: The abnormally low expression of ARHGAP5-AS1 may affect the occurrence and development of breast cancer by promoting the proliferation, migration and invasion of breast cancer cells.