| Objective: To detect the expression level of linc00324 in breast cancer and explore the relationship between linc00324 and the clinical signification of breast cancer,and to explore the role of linc00324 in the proliferation,migration,invasion and apoptosis of breast cancer cell line MDA-MB-231.Method:1.The relationship between linc00324 expression level and clinical parameters was analyzed of using TCGA Database and GEO Database,and its diagnostic and prognostic value was evaluated by ROC cruve.2.Samples of breast cancer tissues were collected from 39 clinical patients in The First Affiliated Hospital of Guangxi Medical University from August 2016 to June 2017.The relative level of linc00324 expression in breast cancer and paracarcinoma tissues were detected by qRT-PCR,and the relationship between the expression level of linc00324 and breast cancer was analyzed by stastical analysis,the clinical factors of patients,including age,sex,TNM stage,PR,ER,Her-2,Ki67,etc.The expression of CTC1 and WT1 in breast cancer tissues was also detected by qRT-PCR and Immunohistochemistry,and the relationship between the expression of linc00324 and CTC1 and WT1 was analyzed.3.A recombinant lentivirus containing linc00324 was constructed,and then transfect human breast cancer cell line MDA-MB-231.The cell proliferation was observed in vitro by CCK-8 assay,and the invasion and migration ability were detected by transwell migration assay and wound healing test,and the apoptosis was evaluated by flow cytometry.Results:1.The expression of linc00324 in breast cancer was analyzed by bioinformatics:(1)The expression of Linc00324 in breast cancer was significantly decreased(p<0.0001),especially in progesterone receptor-negative,estrogen receptor ER-negative or human epidermal growth factor2(Her-2)-negative patients.(2)The expression level of linc00324 was related to histological grade and T stage.(3)The expression of Linc00324 was positively correlated with the expression of CTC1.(4)The area under the ROC curve of linc00324 was 0.722(Se=0.667,Sq=0.60)(p<0.000),and the area under the ROC curve of Ki-67 was 0.722(Se=0.816,Sq=0.70)(p<0.0001).The area under combined ROC curve is 0.928(Se=0.98,Sq=0.88)(p<0.0001),suggesting that the combined detection of Ki-67 and linc00324 has good clinical diagnostic value for breast cancer patients.2.Expression and clinical significance of Linc00324 in breast cancer:(1)The expression of linc00324 was significantly lower in breast cancer tissues,and was related to histological grade(P < 0.05).(2)The expression of WT1 was significantly increased in breast cancer tissues,especially in ER-negative patients,and was negatively correlated with linc00324 expression.(3)The expression of CTC1 in breast cancer tissues was significantly decreased.3.The effect of linc00324 on breast cancer cell biological behavior:(1)The expression of linc00324 in breast cancer cell line MDA-MB-231 was significantly lower than that in normal breast cell line MCF-10A.(2)CCK8results showed that overexpression of linc00324 significantly inhibited the proliferation of MDA-MB-231 cells.(3)Transwell assay and wound healing test showed that the overexpression of linc00324 significantly reduced the ability of cell invasion and migration.(4)FACS analysis showed that overexpression of linc00324 could induce cell apoptosis.(5)Overexpression of linc00324 has no effect on the expression ofβ-catenin.Conclusion: The expression of Linc00324 was significantly decreased in breast cancer tissues,which was significantly correlated with the histological grade.Overexpression of linc00324 inhibits the proliferation,invasion and migration of breast cancer cells and induces apoptosis.These results suggest that linc00324 may plays an important role in the occurrence and development of breast cancer,and may provides a new idea for diagnosis,targeted therapy and prognosis of breast cancer. |
PDF Full Text Request