Tumor-associated neutrophils induce EMT through IL-17a promotes migration and invasion in colon cancer
Objective To investigate the role and mechanism of tumor-associated neutrophils(TANs)in promoting the migration and invasion in colon cancer through interleukin-17a(IL-17a).Methods Neutrophils isolated from healthy donors were stimulated with fresh colon cancer tissue culture supernatants and co-cultured with HCT116 cells as observation group one,anti-IL-17a receptor antibody treatment based on observation group one as observation group two,and the control group was simple HCT116 cells.Migration and invasion assays were used to measure cell migration and invasion capacity,and the EMT markers E-cadherin and Vimentin protein were detected by Western blot.Changes in cell migration ability,changes in cell invasion ability,and cellular E-cadherin and Vimentin protein expression were analyzed and compared among the three groups.Results The number of cell migration was(52.00±4.35)in the control group and(113.67±5.81)in the observation group one.The number of cell migration was significantly increased in the observation group one compared with the control group(t=8.488,P=0.001<0.05).After the addition of neutralizing antibody to block IL-17a,the number of cell migration in the observation group two was(54.33±3.97),which was significantly reduced compared with the observation group one(t=8.457,P=0.001<0.05).The number of invaded cells was(43.00±2.65)in the control group and(95.00±4.04)in the observation group one.The number of invaded cells was significantly increased in the observation group one compared with the control group(t=10.770,P=0.000<0.05).After the addition of neutralizing antibody to block IL-17a,the number of invaded cells in the observation group two was(45.33±3.28),which was significantly reduced compared with the observation group one(t=9.542,P=0.000<0.05).The relative expression of E-cadherin protein in the control group was(0.230±0.020),and that in the observation group one was(0.003±0.004).Compared with the control group,the expression of E-cadherin in the observation group one was significantly decreased(t=11.351,P=0.000<0.05).After the addition of neutralizing antibody to block IL-17a,the relative expression of E-cadherin protein in the observation group two was(0.200±0.003),which was significantly increased compared with that in the observation group one(t=-11.548,P=0.000<0.05).The relative expression of Vimentin in the control group was(0.770±0.030),and that in the observation group one was(0.890±0.060).Compared with the control group,the expression of Vimentin in the observation group one was significantly increased(t=-4.081,P=0.015<0.05).After the addition of neutralizing antibody to block IL-17a,the relative expression of Vimentin in the observation group two was(0.003±0.001),which was significantly lower than that in the observation group one(t=26.566,P=0.000<0.05).Conclusion TANs promote migration and invasion ability by inducing EMT in colon cancer cells through IL-17a.Blocking IL-17a can inhibited the stimulus-inducing effects of TANs.Targeting IL-17a may have the potential to treat colon cancer.
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