Metallothionein 1M inhibits migration and invasion of triple-negative breast cancer cells by regula-ting phosphatidylinositol-3-kinase-protein kinase B/epithelial mesenchymal transformation
Objective To observe the expression of metallothionein 1M(MT1M)in triple-negative breast cancer(TNBC)cells,and study the effect and mechanism of MT1M on TNBC cell migration and inva-sion.Methods Ualcan and Kaplan-Meier Plotter online websites were used to analyze the expression of MT1M in TNBC and normal breast tissues and the relationship between MT1M expression level and the prog-nosis of TNBC patients.TNBC cells(MDA-MB-231 and BT-549)were transfected with MT1M overexpression plasmid as MT1M overexpression group,and TNBC cells transfected with empty plasmid as control group.Western blotting was used to verify the MT1M protein level in the transfected cells of the two groups.The mi-gration and invasion ability of cells was detected by scratch test and Transwell chamber test.Western blotting was used to detect the levels of E-cadherin,Vimentin,phosphatidylinositol-3-kinase(PI3K),protein kinase B(Akt)and phosphorylated protein kinase B(p-Akt)in the two groups.The t-test was used for comparison between the two groups.Results Website data analysis showed that MT1M mRNA was abnormally low in TNBC,and patients with abnormally low MT1M expression had poor prognosis.The protein expression of MT1M in MDA-MB-231 and BT-549 cells in MT1M overexpression group(11.08±1.90,9.00±0.69)was significantly higher than that in control group(1.35±0.52,1.41±0.43,t=8.569,16.220,P＜0.05).The wound healing rate of MDA-MB-231 cells in MT1M overexpression group[12 h(25.46±10.31)％;24 h(44.55±23.65)％]was significantly lower than that in control group[12 h(33.07±9.28)％;24 h(60.68±21.89)％,t=4.426,8.202,P＜0.05].The wound healing rate of BT-549 cells in MT1M overex-pression group[24 h(30.40±11.52)％;48 h(64.01±7.28)％]was significantly lower than that in con-trol group[24 h(57.21±11.98)％;48 h(98.99±0.29)％,t=6.780,8.660,P＜0.05].Transwell as-say results showed that the number of MDA-MB-231 and BT-549 cells passing through(141.67±1.53,173.00±11.27)in the MT1M overexpression groups was significantly less than that in the control group(339.67±61.86,376.00±19.00,t=5.678,17.390,P＜0.05).The expression of E-cadherin protein in MDA-MB-231 and BT-549 cells in MT1M overexpression group(4.80±0.65,4.81±1.20)was significantly higher than that in control group(1.29±0.26,1.41±0.68,t=8.745,4.279,P＜0.05).The expression of Vimentin,PI3K,Akt and p-Akt was 1.25±0.32,1.11±0.13;1.40±0.52,1.52±0.48;1.05±0.07,1.04±0.05 and 1.35±0.51,1.06±0.07,significantly lower than the control group(5.16±1.10,6.35± 0.13;4.52±0.50,3.37±0.55;1.90±0.18,1.98±0.19;4.55±0.68,3.10±0.59,t=5.932,48.25;7.525,4.420;7.614,8.466;6.524,5.961,P＜0.05).Conclusion The expression of MT1M was low in TNBC cells.MT1M may affect the migration and invasion of TNBC cells by regulating PI3K-Akt/epithelial mesenchymal transformation.
Metallothionein 1MTriple negative breast cancerEpithelial mesenchymal trans-formationMigrationInvasion
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