Study On The Role And Mechanism Of BP-1-102 In Regulating The Progression Of Pituitary Adenomas Through Macrophages

Objective This study aims to reveal whether BP-1-102 regulates the progression of pituitary adenomas by macrophages,to provide a new experimental basis for BP-1-102 as an anti-tumor drug and to enrich its drug mechanism.Methods 1.Raw264.7 cells were selected and divided into:（1）M（?）group,DMSO group,M1 group,M1+BP(4×10^-6mol·L^-1)group,M1+BP(6×10^-6mol·L^-1)group,M1+BP(8×10^-6mol·L^-1)group;（2）M（?）group,DMSO group,M2 group,M2+BP(4×10^-6mol·L^-1)group,M2+BP(6×10^-6mol·L^-1)group,M2+BP(8×10^-6mol·L^-1)group.Raw264.7 cells were treated with BP-1-102 for 24h,and then they were induced into M1 type（F4/80+CD86）and M2 type（F4/80+CD206）macrophages.Flow cytometry was used to detect the expression level of F4/80+CD86 or F4/80+CD206 in each group;ELISA and QPCR were used to detect the expression levels of IL-10,VEGF-A and TGF-β1 in each group.2.AtT-20 cells and Raw264.7 cells were selected and divided into PA group,PA+M（?）group,PA+M1 group,PA+M2 group.The macrophages were co-cultured with AtT-20 cells,the invasion and proliferation ability of AtT-20 cells were detected by CCK-8,and the cell cycle and apoptosis level of AtT-20 were detected by flow cytometry.3.AtT-20 cells and Raw264.7 cells were selected and divided into PA+M1 group,PA+M2 group,PA+(M1+BP 8×10^-6mol·L^-1)group,PA+(M2+BP 8×10^-6mol·L^-1)group.Macrophages were intervened with BP-1-102(8×10^-6mol·L^-1)and then co-culture with AtT-20 cells,the invasive and proliferative ability of AtT-20 cells were detected by CCK-8,and the cell cycle and apoptosis levels of AtT-20 were detected by flow cytometry.Results 1.The effect of BP-1-102 on macrophages’polarity and cytokine expression levels（1）Macrophage phenotypes:（1）Comparing the M（?）group with the DMSO group,there was no significant difference in the results.（2）Compared with the M（?）group,the expression of F4/80+CD86 increased in the M1 group;M1 macrophages treated with BP-1-102:With the increase of BP-1-102,the expression of F4/80+CD86 gradually decreased.（3）Compared with M（?）group,the expression of F4/80+CD206 was significantly higher in M2 group;M2 type macrophages treated with BP-1-102:With the increase of BP-1-102,the expression of F4/80+CD206 gradually decreased.（4）Compared with F4/80+CD86,with the increase of BP-1-102,the inhibitory effect on F4/80+CD206 was significantly enhanced.（2）Cytokine levels:（1）Comparing DMSO group with the M（?）group,there was no significant difference in the expression of IL-10,VEGF-A and TGF-β1.（2）Compared with the M（?）group,the expressions of IL-10and TGF-β1 were reduced in the M1 group,and the expression of VEGF-A was increased;M1 type macrophages treated with BP-1-102,with the increase of BP-1-102,the expressions of IL-10 and TGF-β1gradually increased,and the expression of VEGF-A gradually decreased.（3）Compared with the M（?）group,the expressions of IL-10,VEGF-A and TGF-β1 were increased in the M2 group;M2 type macrophages treated with BP-1-102,with the increase of BP-1-102,the expressions of IL-10,VEGF-A and TGF-β1gradually decreased.2.The effect of macrophages on pituitary adenomasCompared with PA group,PA+M（?）group:There was no significant difference in the invasive and proliferative ability,cell cycle and apoptosis level of AtT-20 cell:AtT-20 cell;PA+M1 group:The invasive and proliferative ability of AtT-20 cell weakened,the proportion of cells in G0/G1 phase increased,and the degree of apoptosis increased;PA+M2 group:The invasive and proliferative ability of AtT-20 cell increased,the proportion of cells in G0/G1 phase decreased,and the degree of apoptosis decreased.3.The effect of macrophages treated by BP-1-102 on pituitary adenomasCompared with PA+M1 group,PA+(M1+BP 8×10^-6mol·L^-1)group:There was no significant difference in the proliferative ability of AtT-20 cells,the invasive ability of these cells was improved,the proportion of cells in S phase+G2/M phase increased,and the degree of apoptosis decreased.Compared with PA+M2group,PA+(M2+BP 8×10^-6mol·L^-1)group:The invasive and proliferative ability of AtT-20 cells decreased,the proportion of cells in S phase+G2/M phase decreased,and the degree of apoptosis increased.Conclusion BP-1-102 regulates the polarity of macrophages and the expression of cytokines,thereby regulating the invasion,proliferation,cycle and apoptosis of pituitary adenoma cells.