Effect of type X collagen alpha 1 chain protein on proliferation,migration and angiogenesis of triple-negative breast cancer cells
Objective To explore the expression and potential biological role and mechanism of type X collagen alpha 1 chain protein(COL1OA1)in triple-negative breast cancer(TNBC),so as to provide new ideas for the diagnosis and treatment of TNBC.Methods Ualcan and Kaplan-Meier were used to investigate the expression of COL10A1.Small interfering RNA(siRNA)and the recombinant overexpressed plasmid GV703-COL10A1 were transfected to get the cell models(Si-COL10A1,Si-NC and OE-COL10A1,NC)of MDA-MB-231 and BT-549.The proliferation and migration ability of TNBC cells was investigated by cell clone formation assay,cell counting kit-8(CCK-8)assay,scratch assay,and cell migration assay,and the tubule formation assay was used to examine the vascular formation ability of human umbilical vein endothelial cells(HUVECs).The comparisons between groups were carried out by t-test.Results Biogenic analysis and Western blotting results showed that COL10A1 was highly expressed in TNBC and high expression COL10A1 was associated with worse overall survival(OS)and progression free survival(RFS).After trans-fection with siRNA or recombinant COL10A1 overexpressed plasmid,Western blotting results showed that COL10A1 protein was significantly knocked down or enhanced in the cells,respectively.The cell clone for-mation experiment showed that the clone formation rates of MDA-MB-231 and BT-549 cells in COL10A1 knockdown group[(10.47±0.91)％,(11.10±1.35)％]were significantly lower than those in the control group[(16.77±2.33)％,(18.43±2.40)％,t=4.373,4.609,P＜0.05].After overexpression of COL10A1,the clone formation rates of MDA-MB-231 and BT-549 cells[(21.50±0.62)％,(27.83± 3.72)％]were significantly higher than those in the control group[(15.23±2.79)％,(19.40±1.47)％,t=3.792,3.648,P＜0.05].The scratch healing rates of MDA-MB-231 and BT-549 cells in the COL10A1 knockdown group[(17.00±1.07)％,(15.38±0.89)％]were significantly lower than those in the control group[(30.58±1.88)％,(23.78±1.58)％,t=6.284,4.636,P＜0.01].The scratch healing rates of MDA-MB-231 and BT-549 cells in the COL10A1 overexpression group[(47.40±3.09)％,(41.26± 4.33)％]were higher than those in the control group[(34.48±2.03)％,(21.80±1.03)％,t=3.491,4.737,P＜0.01].In the cell migration experiment,the number of MDA-MB-231 and BT-549 cells entering the lower chamber[(151.70±33.25),(76.67±10.41)cells]in the COL10A1 knockdown group was lower than that in the control group[(378.00±26.51),(303.70±15.50)cells,t=9.219,21.060,P＜0.01].After overexpression of COL10A1,the number of of MDA-MB-231 and BT-549 cells entering the lower com-partment[(1 519.00±144.10),(551.00±61.65)cells]was greater than that in the control group[(426.30±46.07),(288.30±27.57)cells,t=12.510,6.736,P＜0.01].HUVECs cultured in condi-tioned medium with COL10A1 overexpression cells significantly increased the number of junctions and total segments length of cell tubulogenesis compared with the control group(P＜0.01).Conclusion COL10A1 is highly expressed in TNBC.COL10A1 can enhance the proliferation and migration ability of MDA-MB-231 and BT-549 cells,and promote angiogenesis.
Triple-negative breast cancerType X collagen alpha 1 chain proteinProlifera-tionMigrationAngiogenesis
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