Effects of macrophages under the influence of tuberculosis antigen Ag85 on the proliferation and apoptosis of Hodgkin lymphoma KM-H2 cells / 白血病·淋巴瘤

ABSTRACT

Objective:To explore the effects of macrophages after influenced by tuberculosis antigen Ag85 on the proliferation and apoptosis of Hodgkin lymphoma cells, and to discuss the possible role of tuberculosis infection in the progression of Hodgkin lymphoma.Methods:The indirect co-culture system between Hodgkin lymphoma cell line KM-H2 and human monocytic leukemia cell line THP-1 (simulated macrophage) was established by using Transwell nesting. KM-H2 cells were cultured as KM-H2 group alone, KM-H2 cells interfered with Ag85 were taken as KM-H2+Ag85 group, and KM-H2 cells co-cultured with THP-1 cells were taken as KM-H2+THP-1 group. The co-culture system of KM-H2 cells and THP-1 cells interfered by Ag85 was taken as KM-H2+THP-1+Ag85 group. The proliferation of KM-H2 cells in each group was detected by using CCK-8 assay, and the growth curve was drawn. The apoptosis of cells in each group was detected by using flow cytometry. The mRNA expression levels of p53, c-myc, bcl-2 and vascular endothelial growth factor receptor 3 (VEGFR3) in each group were detected by using quantitative real-time fluorescence polymerase chain reaction (qRT-PCR). The expressions of bax and bcl-2 proteins were detected by using Western blotting.Results:The cell proliferation ability of KM-H2+Ag85 group was higher than that of KM-H2 group (all P = 0.001) after 24 and 48 h culture, but the cell proliferation ability of KM-H2+THP-1 group was lower than that of KM-H2 group after 24 h, 48 h and 72 h culture (all P < 0.05). The cell proliferation ability of KM-H2+THP-1+Ag85 group was lower than that of KM-H2 group after 48 h and 72 h culture (all P < 0.05), but the cell proliferation ability of KM-H2+THP-1+Ag85 group was enhanced after 24 h and 48 h culture compared with KM-H2+THP-1 group, and there was no statistically significant difference between the two groups after 72 h culture ( P > 0.05). The apoptosis rate of KM-H2+Ag85 group was lower than that of KM-H2 group [(0.92±0.80)% vs. (6.02±1.63)%, P < 0.001], and the apoptosis rate of KM-H2+THP-1 group [(8.57±0.57)%] was higher than that of KM-H2 group ( P < 0.05). The apoptosis rate [(0.60±0.13)%] in KM-H2+THP-1+Ag85 group was lower than that in KM-H2+THP-1 group ( P < 0.001). The relative expression of bcl-2 and VEGFR3 mRNA in KM-H2+Ag85 group was higher than that in KM-H2 group ( P = 0.018, P = 0.017), while the relative expression of c-myc mRNA in KM-H2+Ag85 group was lower than that in KM-H2 group ( P = 0.016), and there was no statistically significant difference of p53 mRNA relative expression level between the both groups ( P > 0.05).The relative expression of p53 mRNA in KM-H2+THP-1+Ag85 group was lower than that in KM-H2+THP-1 group ( P = 0.048), while the relative expressions of bcl-2 and VEGFR3 mRNA in KM-H2+THP-1+Ag85 group were higher than those in KM-H2+ THP-1 group ( P = 0.016; P = 0.021). The expression of bax protein in KM-H2+Ag85 group was lower than that in KM-H2 group ( P = 0.019), and bcl-2 protein was more than that in KM-H2 group ( P = 0.001). The expression of bax protein in KM-H2+THP-1+Ag85 group was lower than that in KM-H2+THP-1 group ( P = 0.011), but there was no statistically significant difference in the expression of bcl-2 protein between the two groups ( P > 0.05). Conclusions:Tuberculosis antigen Ag85 may inhibit the apoptosis of Hodgkin lymphoma KM-H2 cells and enhance the proliferative activity by affecting the function of macrophages.