LncRNA A2M-AS1 Promotes Proliferation,Invasion And Migration Of Breast Cancer And Is Associated With Poor Prognosis

ObjectiveThe purpose of this study was to investigate the clinical value of long non-coding RNA(lncRNA)A2M-AS1 in breast cancer,and to investigate its function and potential molecular mechanisms in breast cancer cells.Methods1?The raw data of breast cancer gene sequencing was downloaded from the Gene expression omnibus(GEO)database,and all differential expressed mRNAs and lncRNAs were screened by R language.The clinical significance of differential expressed lncRNA,A2M-AS1,was analyzed.2?The real-time fluorescence quantitative reverse transcription PCR(qRT-PCR)was used to detect the expression of A2M-AS1 in 96 human breast cancer tissues and 64 adjacent cancer tissues.3?According to the average expression of A2M-AS1,A2M-AS1 was divided into high expression group and low expression group.Kaplan-meier analysis showed the relationship between A2M-AS1 expression and clinicopathological features and prognosis of breast cancer.4.The effect of A2M-AS1 on the proliferation of breast cancer cells was detected by CCK-8 assay.5.Transwell and scratch assays were used to detect the effect of A2M-AS1 on invasion and migration of breast cancer cells.6.Knockdown and over-expressed A2M-AS1,qRT-PCR and Western blot(WB)assays were used to detect the CD2 and SELL in mRNA and protein levels.Result1.In GSE42568 and GSE65194,we found that A2M-AS1 is significantly higher in human breast cancer tissues than in adjacent normal tissues,and its high expression is associated with estrogen receptor(ER)negative,basal-like and human epidermal growth factor receptor-2(HER-2)positive breast cancer,as well as shorter recurrence-free survival and shorter metastatic-free survival.2.The expression of A2M-AS1 was verified by qRT-PCR in 96 breast cancer tissues,collected from the Affiliated Hospital of Jiangnan University.The result showed that A2M-AS1 is significantly higher in human breast cancer tissues than in adjacent normal tissues.A2M-AS1 is associated with ER-negative patients,shorter recurrence-free survival and shorter metastatic-free survival.3.Knockdown of A2M-AS1 in breast cancer cell lines ZR-75-30 and BT-549,which high expression of A2M-AS1,significantly inhibited the proliferation,invasion and migration of these breast cancer cells.Over-expression of A2M-AS1 in MDA-MB-231 and Hs578 T in breast cancer cell lines,which low expression of A2M-AS1,significantly promoted proliferation,invasion and migration of these breast cancer cells.4.Bio-informatics analysis of genes regulated by A2M-AS1,then verified by qRT-PCR and Western blot,we found that A2M-AS1 may affect the invasion and migration of breast cancer by regulating CD2 and SELL.Conclusion1.A2M-AS1 is up-regulated in breast cancer,and its high expression is associated with malignant pathological features and poor prognosis.2.A2M-AS1 is first discovered to promote proliferation,invasion and migration in breast cancer cells.3.A2M-AS1 might promote the invasion and migration of breast cancer cells by up-regulating CD2 and SELL.We have discovered and identified a new breast cancer carcinogenic lncRNA A2M-AS1,and elucidated its potential molecular mechanism,which can be used as a prognosis biomarker and therapeutic targets for breast cancer,laying a theoretical and experimental basis for the prevention and treatment of breast cancer.