title: [Inducing apoptosis and reversal effect of nilotinib in combination with tetrandrine on multidrug resistance of K562/A02 cell line].
abstract: This study was aimed to investigate the relevance of nilotinib in combination with tetrandrine (Tet) on reversing multidrug resistance and inducing apoptosis of K562/A02 cell line and its mechanism. Methyl-thiazol tetrazolium (MTT) assay was employed to examine the pharmacological effect of nilotinib or Tet alone on K562/A02 cell line, the IC(50) of daunorubicin (DNR) on K562/A02 cell line treated with nilotinib and Tet was calculated; the flow cytometry (FCM) was employed to detect the apoptosis rate of K562/A02. The expression of bax/survivin mRNA was determined by RT-PCR, and the expression of bax/survivin protein was assayed by Western blot. The results showed that after being treated by 5 nmol/L nilotinib or 1.0 µml/L Tet for 48 hours, IC(50) of DNR to K562/A02 was 5.71 ± 0.72 mg/L or 6.52 ± 0.43 mg/L, respectively, while in their combined treatment, IC(50) decreased to 3.12 ± 0.13 mg/L. Nilotinib or Tet alone could increase DNR-inducing apoptosis rate of K562/A02 cell, while the apoptosis rate of K562/A02 increased remarkably in combination treatment of nilotinib with Tet. After being treated with 5 nmol/L nilotinib or 1.0 µml/L Tet alone for 48 hours, the expressions of bax mRNA and BAX protein was up-regulated, while both effects were more obvious in combination treatment of nilotinib with Tet. Treatment with 5 nmol/L nilotinib or 1.0 µmol/L Tet alone for 48 hours down-regulated the expression of survivin mRNA and its protein, while treatment of nilotinib in combination with Tet had more significant effect on down-regulation of their expression. It is concluded that the K562/A02 cells are resistant to DNR, nilotinib or Tet alone both can partially reverse resistance of K562/A02 cells to DNR, increase the apoptosis rate of K562/A02 cells. Combination of nilotinib with Tet shows obvious synergistic action, mechanism of which may associate with up-regulation of bax mRNA and BAX protein expressions and down-regulation of survivin mRNA and its protein expressions.
authors.affiliation: Department of Hematolagy, Zhongda Hospital, Southeast University Medical College, Nanjing 210009, Jiangsu Province, China.
authors.forename: Ting-Yun
authors.initials: TY
authors.lastname: Cui
authors.forename: Bao-An
authors.initials: BA
authors.lastname: Chen
authors.forename: Jia-Hua
authors.initials: JH
authors.lastname: Ding
authors.forename: Chong
authors.initials: C
authors.lastname: Gao
authors.forename: Jian
authors.initials: J
authors.lastname: Cheng
authors.forename: Wen
authors.initials: W
authors.lastname: Bao
authors.forename: Yue-Jiao
authors.initials: YJ
authors.lastname: Zhong
authors.forename: Xue-Yun
authors.initials: XY
authors.lastname: Shan
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
authors.forename: Guo-Hua
authors.initials: GH
authors.lastname: Xia
authors.forename: Anita
authors.initials: A
authors.lastname: Schmitt
authors.forename: Michael
authors.initials: M
authors.lastname: Schmitt
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D044182:Benzylisoquinolines / Q000008:administration & dosage / Q000494:pharmacology*; D003630:Daunorubicin / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D015973:Gene Expression Regulation, Leukemic; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics; D020014:K562 Cells; D011743:Pyrimidines / Q000008:administration & dosage / Q000494:pharmacology*; D000077022:Survivin; D051028:bcl-2-Associated X Protein / Q000235:genetics
chemical_list: C494826:BAX protein, human; C107489:BIRC5 protein, human; D044182:Benzylisoquinolines; D051034:Inhibitor of Apoptosis Proteins; D011743:Pyrimidines; D000077022:Survivin; D051028:bcl-2-Associated X Protein; C009438:tetrandrine; C498826:nilotinib; D003630:Daunorubicin