The Role Of SMOC2 In The Proliferation,Migration And Invasion Of Triple Negative Breast Cancer MDA-MB-231 Cells

Part ⅠConstruction of SMOC2 Gene Interference or Overexpressionstable MDA-MB-231 Cell LineObjective:To construct a lentiviral-mediated SMOC2 gene knockdown stable cell line and negative control cell line of MDA-MB-231 cells,construction of lentiviral-mediated stable cell line of SMOC2 gene overexpression in MDA-MB-231 cells and Negative control cell line.Method:1.shSMOC2 vector and its negative control vector shNC were purchased from Shanghai Jima Co.,2.Packaging viral vector with 293T cells;3.Infecting MDA-MB-231 cells with virus.4.Continuous killing of cells with puromycin.5.Detection efficiency using Western blot and RT-qPCR.Results:1.Compared with shNC empty vector group(1.00±0.28),MDA-MB-231 cells infected with shSMOC2 interference vector(0.21±0.0141),the mRNA level and protein level of SMOC2 were significantly decreased(p<0.01).Conclusions:Obtaining a lentivirus-mediated stable cell line of SMOC2 gene knockdown of MDA-MB-231 cells and its negative control cell line,Obtaining a lentiviral-mediated stable cell line overexpressing the SMOC2 gene of MDA-MB-231 cells and a negative control cell line thereof.Part ⅡThe Role of SMOC2 in MDA-MB-231 Cell Proliferation,Migrationand InvasionObjective:To study the role of SMOC2 in the proliferation,migration and invasion of MDA-MB-231 cells.Methods:Detection of cell proliferation,migration and invasion by CCK-8,EdU,monoclonal formation,flow cytometry,scratch and Transwell assayResults:1.CCK-8 experiment:the light absorption value of the knockdown group was significantly higher than that of the control empty vector group(p<0.001);the light absorption value of each time point of the overexpression group was lower than that of the control empty vector group.Among them,there were significant differences between the 48 hours and 72 hours(p<0.001).2.EdU experiment:Compared with the control empty vector group(0.5070±0.0115),the insertion ratio of EdU increased in the SMOC2 interference group(0.5597±0.0106)(p<0.05);the EdU insertion of the SMOC2 overexpression group(0.4445±0.0108)The ratio was lower than that of the control empty vector group(0.5056 ± 0.0143)(p<0.05).3.Monoclonal formation assay:Compared with the control empty vector group(46.2 ±5.250),the number of clone formation was significantly increased in the SMOC2 interference group(90.2±24.503)(p<0.01).4.Flow cytometry to detect cell cycle:Compared with the control empty vector group(0.2640±0.0161),the S phase of the SMOC2 overexpression group(0.4867±0.01 3 8)was significantly blocked(p<0.001).5.Transwell assay migration:Compared with the control empty vector group(1720±510),the number of migrated cells in the SMOC2 knockdown group was significantly decreased(p<0.001);the SMOC2 overexpression group had significantly more migrated cells than the control empty vector.Group(45.2 ± 9.2356)(p<0.001)· 6.Compared with the control empty vector group,the number of invasive cells in the SMOC2 knockdown group was significantly decreased(p<0.001);the number of invasive cells in the SMOC2 overexpression group was significantly higher than that in the control empty vector group(p<0.05).Conclusion:The function of SMOC2 is mainly to inhibit proliferation,promote migration and invasion.