1.Effect of Qiling Baitouweng Tang on Proliferation and Apoptosis in Diffuse Large B-cell Lymphoma Through JAK2/STAT3 Signaling Pathway
Ning LIU ; Xinzhuo ZHAN ; Hui YU ; Xiaoli CHEN ; Xiangtu KONG ; Haiwen NI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(13):10-19
ObjectiveTo investigate the effect of Qiling Baitouweng Tang (QLBTWT) on proliferation and apoptosis, Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway and interleukin-10 (IL-10) in diffuse large B-cell lymphoma (DLBCL). MethodWith human DLBCL cells OCI-LY10 and U2932 as research objects, cell proliferation was detected by cell counting kit-8 (CCK-8) assay. After treatment with 0, 4.6, 9.3, 18.7, 37.5, 75, 150 mg·L-1 QLBTWT for 24 h, the half-inhibitory concentration (IC50) of OCL-LY10 and U2932 cells was calculated to be 9.33, 16.13 mg·L-1, respectively, based on which, 9.5, 19, 38 mg·L-1 QLBTWT were selected for subsequent experiments. After 0, 9.5, 19, 38 mg·L-1 QLBTWT treatment for 24 h, the zymogen activities of Caspase-3, Caspase-8 and Caspase-9 in OCI-LY10 and U2932 cells were detected using corresponding activity assay kits (colorimetric), and the IL-10 expression was detected by enzyme-linked immuno sorbent assay (ELISA). The apoptosis rate and cell cycle of OCI-LY10 and U2932 cells treated with different concentrations of QLBTWT for 24 h were detected by flow cytometry. The expressions of apoptosis-related proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved poly adenosine diphosphate ribose polymerase (cleaved PARP), cleaved Caspase-3], JAK2, STAT3, phospho-JAK2 (p-JAK2), phospho-STAT3 (p-STAT3) pathway proteins, and c-Myc protein in OCL-LY10 and U2932 cells after 24 h treatment with 0, 9.5, 19, 38 mg·L-1 QLBTWT were all tested by Western blot. ResultAfter QLBTWT treatment on OCI-LY10 and U2932 cells for 24 h, cell proliferation was inhibited in each QLBTWT group compared with that in the control group (P<0.05, P<0.01). The zymogens of Caspase-3, Caspase-8 and Caspase-9 were activated (P<0.01), and there was an increase in cell apoptosis (P<0.05, P<0.01) and cell cycle arrest at Gap phase1 (G1) phase in 9.5, 19 and 38 mg·L-1 QLBTWT group (P<0.05, P<0.01). After 9.5, 19 and 38 mg·L-1 QLBTWT treatment on OCI-LY10 and U2932 cells for 24 h, the expressions of Bcl-2, p-JAK2 and p-STAT3 proteins were decreased (P<0.01), and the expressions of Bax, cleaved PARP and cleaved Caspase-3 proteins were increased (P<0.01), but no significant change was observed in the expressions of JAK2 and STAT3 proteins. Compared with the conditions in the control group, the expressions of c-Myc, p-JAK2, and p-STAT3 proteins were down-regulated in 19 mg·L-1 QLBTWT group and 19 mg·L-1 QLBTWT+10 μg·L-1 IL-10 group (P<0.05, P<0.01), and up-regulated in 10 μg·L-1 IL-10 group (P<0.05, P<0.01), while there was no difference in JAK2/STAT3 proteins. ConclusionQLBTWT can inhibit proliferation and induce apoptosis of human DLBCL cells OCI-LY10 and U2932, and the potential mechanism may be related to the regulation of JAK2/STAT3 signaling pathway.