title: [Effect of PCI-32765 and bortezomib on proliferation and apoptosis of B-cell tumor cell lines and its mechanisms].
abstract: This study was aimed to investigate the effect of Btk inhibitor PCI-32765 and the proteasome inhibitor bortezomib on Raji and Ramos cell proliferation, apoptosis, and its mechanisms. Raji and Ramos cells were treated with PCI-32765 and bortezomib alone and/or their combination. The cell proliferation and apoptosis were detected by CCK-8 and flow cytometry respectively, the expression level of Btk, NFκB, c-IAP1, Bcl-xL and caspase-3 protein were measured by Western blot. The results indicated that: (1) after Raji and Ramos cells were treated with PCI-32765 (0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0 µmol/L) alone and bortezomib (10, 20, 30, 40, 50, 60, 80 nmol/L) alone and their combination for 48 h, the cell proliferation and vitality were inhibited in a dose-dependent manner and both had synergistic effect; (2) Raji and Ramos cells were treated with PCI-32765 (2.0 µmol/L) and bortezomib (20 nmol/L) alone and their combination for 8, 12, 24, 36, 48 and 72 h, the cell proliferation and vitality were inhibited in a time-dependent manner, the two drugs displayed a synergistic effects; (3) the Raji and Ramos cells were treated with PCI-32765 (2.0 µmol/L) and bortezomib (20 nmol/L) alone and their combination for 48 h, all these treatments could induce significant apoptosis of Raji and Ramos cells.In Raji cell experiment, the cell apoptosis rate in the control group, PCI-32765 group, bortezomib group and PCI-32765 and bortezomib combination group were 10.34 ± 0.53%, 24.26 ± 0.91%, 43.66 ± 1.08% and 74.06 ± 0.72% respectively, and the differences was statistically significant among the different groups (P < 0.05). In Ramos cell experiment, the cell apoptosis rate in the control group, PCI-32765 group, bortezomib group and PCI-32765 and bortezomib combination group are 15.16 ± 1.49%, 71.36 ± 0.82%, 75.32 ± 2.36% and 84.30 ± 0.91% respectively, the differences was statistically significant among the different groups (P < 0.05); (4) PCI-32765 and bortezomib could inhibit the expression level of intracellular Btk, NFκB, Bcl-xl and c-IAP1 proteins, but up-regulate the expression level of caspase-3. It is concluded that PCI-32765 and bortezomib can synergistically inhibit the proliferation and induce apoptosis of Raji and Ramos cells, the mechanism may be associated with inhibition of Btk and NFκB activity, down-regulation of anti-apoptotic proteins expression, such as Bcl-xl and c-IAP1, and increase of caspase-3 expression.
authors.affiliation: Department of Hematology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an 223300, Jiangsu Province, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Deng
authors.forename: Shan-Dong
authors.initials: SD
authors.lastname: Tao
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.forename: Zheng-Mei
authors.initials: ZM
authors.lastname: He
authors.forename: Yue
authors.initials: Y
authors.lastname: Chen
authors.forename: Zhi-Kui
authors.initials: ZK
authors.lastname: Deng
authors.forename: Yuan-Yuan
authors.initials: YY
authors.lastname: Li
authors.forename: Liang
authors.initials: L
authors.lastname: Yu
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives; D000077329:Agammaglobulinaemia Tyrosine Kinase; D017209:Apoptosis / Q000187:drug effects*; D001897:Boronic Acids / Q000494:pharmacology*; D000069286:Bortezomib; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D015973:Gene Expression Regulation, Leukemic; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D010880:Piperidines; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011719:Pyrazines / Q000494:pharmacology*; D011720:Pyrazoles / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D051020:bcl-X Protein / Q000378:metabolism
chemical_list: C494811:BCL2L1 protein, human; D001897:Boronic Acids; D051034:Inhibitor of Apoptosis Proteins; D016328:NF-kappa B; D010880:Piperidines; D011719:Pyrazines; D011720:Pyrazoles; D011743:Pyrimidines; D051020:bcl-X Protein; C551803:ibrutinib; D000069286:Bortezomib; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; C000625949:BTK protein, human; C505407:CASP3 protein, human; D053148:Caspase 3; D000225:Adenine