| OBJECTIVE: To investigate the expression and clinical significance of H19,HOTAIR,LSINCT5,MALAT-1 and GAS5 in breast cancer tissue and plasma;preliminary clarify the effect of MALAT-1 on biological behavior of triple-negative breast cancer MDA-MB-231 cells.METHODS: 1.Series of lnc RNA levels were measured in the 60 pairs of human breast cancer tumor and matched para-carcinoma tissues using quantitative real-time PCR,lnc RNA levels in the plasma were measured in 46 patients with benign breast disease,60 patients with breast cancer and 82 healthy adults using quantitative real-time PCR.The correlations of lnc RNA in breast cancer tissue and plasma were analyzed.The correlations of plasma lnc RNA level with the clinicopathological features of the breast cancer patients were analyzed.2.The content of CA125 and CA153 in plasma was detected by electrochemiluminescence,a multivariate logistic regression model was established to analyze the diagnostic efficacy of plasma lnc RNA,CA125 and CA153.3.The expression of MALAT-1 in human breast cancer cell lines MCF-7,MCF-10 A,MDA-MB-231 was analyzed using quantitative real-time PCR.According to the targeted gene MALAT-1,we constructed MALAT-1-si RNA and then transfected it into triple-negative breast cancer MDA-MB-231 cells to knock down the expression of MALAT-1.MTT assay was used to detect cell proliferation,cell migration and invasion were assessed by wound healing assay and transwell assay.RESULTS: 1.Quantitative real-time PCR showed that compared with the matched para-carcinoma tissues,the expressions of H19,HOTAIR,LSINCT5 and MALAT-1 in breast cancer tissue were increased(all P<0.001),the expression of GAS5 was decreased(P<0.001).The lnc RNA levels in the plasma had no statistical difference between benign and healthy groups(all P>0.05).The expressions of H19,HOTAIR and MALAT-1 in the plasma of breast cancer patients were significantly higher than that in the healthy group(all P<0.001),plasma GAS5 level was lower than that in the healthy group(P<0.001),plasma LSINCT5 level has no statistical difference between breast cancer and healthy groups(P>0.05).The expressions of H19,HOTAIR,MALAT-1,GAS5 in breast cancer tissue and plasma were positively correlated(respective rs: 0.44,0.42,0.70,0.61,all P<0.05).Plasma H19 level was significantly correlated with ER and HER-2 level(P<0.05),plasma HOTAIR level was significantly correlated with ER level(P<0.001)and lymph node metastasis(P<0.05),plasma MALAT-1 level was significantly correlated with triple-negative breast cancer(P<0.05)and lymph node metastasis(P<0.001),while plasma LSINCT5 and GAS5 level were not correlated with clinicopathological features(P>0.05).2.The area under the ROC curve of plasma H19,HOTAIR and MALAT-1 was 0.848,0.917 and 0.906,which was much higher than those of CA125(AUC=0.618,P<0.001)and CA153(AUC=0.623,P<0.001).3.RT-PCR showed the expressed of MALAT-1 in cell lines MDA-MB-231 was higher than in MCF-7(P<0.05)and MCF-10A(P<0.01);24hours after transfecting,the expression level of MALAT-1 in interference group was significantly lower than negative control group(P<0.001),the result of MTT assay indicated that cell proliferative activity was lower than that of the control group(P<0.05),wound healing test and transwell showed that the migration ability and the invasive rate was lower than that of the control group(all P<0.05).CONCLUSION: 1.The expressions of plasma H19,HOTAIR,MALAT-1 and GAS5 were significantly different in breast cancer,the diagnostic efficacy of plasma H19,HOTAIR and MALAT-1 was satisfactory for breast cancer,the expressions of plasma H19,HOTAIR and MALAT-1 were significantly correlated with clinicopathological features,they could be the potential biomarker for breast cancer diagnosis.2.Plasma MALAT-1 level was significantly correlated with triple-negative breast cancer,downregulating the expression of MALAT-1 in MDA-MB-231 cells inhibited the proliferation of breast cance cells,and decreased the ability of invasion and migration,MALAT-1 could be the potential diagnostic biomarker and new therapeutic target for triple-negative breast cancer. |
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