Journal List > Korean J Lab Med > v.27(4) > 1011399

Do, Oh, Song, Chung, Kang, and Lee: Treatment Outcome of Multidrug Resistance Related mRNA Expression and c-Jun-N-Terminal Kinase Activity in Patients with Acute Myeloid Leukemia

Abstract

Background

The multidrug resistance (mdr1), multidrug resistance associated protein (mrp1), and glutathione-s-transferase (gst) π genes have been associated with treatment failure in acute myeloid leukemia (AML). c-jun N-terminal kinase (JNK) activity is increased in response to chemotherapeutic agent.

Methods

To investigate the significance of multidrug resistance (mdr) parameters and JNK activity, bone marrow or peripheral blood cells from 52 patients with AML were analyzed. RT-PCR was performed for mdr1, mrp1, and gst π gene expression. JNK expression and activity were measured using an immunoenzymatic kinase assay and a western blot method.

Results

High level expression of mdr1, mrp1, and gst π mRNA was observed in 38.5%, 48.1% and 54.3% of AML cases, respectively. The remission rate was significantly low in cases with an older age (>55 yr), a high WBC count, poor chromosomal abnormalities, a high level expression of mdr1 and mrp1. The WBC count and mdr1 mRNA expression were independent predictors for the outcome to induction chemotherapy. There was a shorter duration of overall survival in the patients with an older age, a high WBC count, chromosome aberrations, high level expressions of mdr1 and mrp1 mRNA, and JNK activation. The patient's age, WBC count and chromosomal abnormalities were independent predictors for overall survivals. The majority (28/30) of AML cases did not show any levels of JNK activation except for two cases, which were associated with an extremely high WBC count, chromosomal aberration, high level expressions of mdr1, mrp1 and gst π mRNA, and treatment resistance.

Conclusions

These data indicate the influences of mdr1 and mrp1 mRNA expression on the clinical outcome of AML to induction chemotherapy. But it will be necessary to investigate further whether blast cells of AML resistant to chemotherapy retain the capacity to activate JNK, and relate to MDR parameters.

References

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Fig. 1.
RT-PCR results for mdr1, mrp1, gst π, and β2-microglobulin mRNA (internal control) with KG1 and CCRF-CEM T-cell lines as mdr1 mRNA positive and negative control and normal bone marrow (A). Expression levels of mdr1, mrp1 and gstπ mRNA using RT-PCR in patients with acute myeloid leukemia (B).
kjlm-27-229f1.tif
Fig. 2.
Immunoblotting of cell lysates of AML patients to analyse c-Jun-N-terminal kinase (JNK) activity and β-actin expression, and JNK activity (phospho c-Jun). JNK activity was undetectable in leukemic samples except 2 cases (middle). P, K562; N, normal bone marrow. Total JNK, 46, 54 kDa; phospho c-Jun, 35 kDa; β-actin; 42 kDa.
kjlm-27-229f2.tif
Table 1.
Clinical and laboratory parameters of patients with acute myeloid leukemia
Parameters  
Male/female (number) 26/26
Age (yr) 44.0* (17–68)
Hemoglobin (g/dL) 8.0 (3.9–15.0)
WBC (×109/L) 32.2 (0.8–206)
Platelets (×109/L) 42.0 (8.0–201.0)
Blasts in blood (%) 62.5 (3.0–97.0)
FAB classification  
M0 3
M1 16
M2 13
M3 11
M4 6
M5 3
Chromosome abnormality  
Favorable 21
Intermediate 26
Unfavorable 5
Treatment response  
CR (%) 63.5 (33/52)

* median;

the parenthesis means the range;

AML patients were grouped into three different risk categories: t(8;21), inv(16), t(15;17) as favorable; −5/−7, 11q23 abnormalities, inv(3q), t(6:9), complex karyotypes as unfavorable. All other aberration and normal karyotypes were regarded as intermediate.

Abbreviation: CR, complete remission.

Abbreviations: mdr1, multidrug resistance; mrp1, multidrug resistance associated protein; gst, glutathione-s-transferase.

Table 2.
Correlation among mdr1, mrp1 and gst π mRNA expression
  mrp1 gstπ
mdr1 0.651 (0.000)* −0.031 (0.839)
mrp1   0.071 (0.641)

* P<0.05.

Abbreviations: See Fig. 1.

Table 3.
Mdr1, mrp1, gstπ mRNA expression and clinical and laboratory findings in patients with acute myeloid leukemia
Parameters mdr1+ (%) P mrp1+ (%) P gst π+ (%) P
Positive N 20/52 (38.5)   25/52 (48.1)   25/46 (54.3)  
Age (yrs)            
<55 11/36 (30.6) (0.122) 14/36 (38.9) (0.071) 18/33 (54.5) (1.000)
55 9/16 (56.3)   11/16 (68.8)   7/13 (53.8)  
Sex            
Male 11/26 (42.3)   11/26 (42.3) (0.579) 15/23 (65.2) (0.236)
Female 9/26 (34.6)   14/26 (53.8)   10/23 (43.5)  
WBC (/L)            
<5 × 109 11/28 (39.3) (0.776) 11/28 (39.3) (0.254) 14/23 (60.9) (0.762)
≥5 × 109 9/22 (40.9)   13/22 (59.1)   11/21 (52.4)  
CD34 (%)            
<20 8/22 (36.4) (1.000) 13/22 (59.1) (0.578) 10/22 (45.5) (0.375)
≥20 12/30 (40.0)   12/30 (40.0)   15/24 (62.5)  
FAB            
M0 2/3 (66.7) (0.123) 1/3 (33.3) (0.154) 2/3 (66.7) (0.228)
M1 5/16 (31.3)   9/16 (56.3)   10/13 (76.9)  
M2 8/13 (61.5)   7/13 (53.8)   5/11 (45.5)  
M3 1/11 (9.1)   2/11 (18.2)   5/10 (50.0)  
M4 3/5 (60.0)   3/5 (60.0)   3/6 (50.0)  
M5 1/3 (33.3)   3/3 (100)   3/3 (100)  
Chromosome            
Good 6/21 (28.6) (0.105) 7/21 (33.3) (0.121) 10/19 (52.6) (0.469)
Intermediate 10/26 (38.5)   14/26 (53.8)   11/22 (50.0)  
Poor 4/5 (80.0)   4/5 (80.0)   4/5 (80.0)  
Treatment            
Response 7/33 (21.2) (0.001*) 12/33 (36.4) (0.043*) 15/29 (51.7) (0.762)
Failure 13/19 (68.4)   13/19 (68.4)   10/17 (58.8)  

* P<0.05; +, expression or overexpression.

Abbreviations: See Fig. 1.

Table 4.
Chemotherapeutic response and overall survivals of AML patients according to clinical findings, and mdr1, mrp1, gst π mRNA and JNK activity
Parameters N of cases Chemotherapy
Overall survival
Remission (%) P Median±SE (months) P
Age          
<55 36 27 (75.0) 0.014* 16.0±3.3 0.001*
≥55 16 6 (37.5)   2.0±3.0  
Sex          
Male 26 15 (57.7) 0.565 60.0±3.1 0.117
Female 26 18 (69.2)   16.0±6.9  
WBC (/L)          
<5×109 28 23 (82.1) 0.004* 17.0±1.8 0.004*
≥5× 109 22 9 (40.9)   3.0±0.5  
CD34 (%)          
<20 24 14 (58.3) 0.568 21.0±3.5 0.129
≥20 28 19 (67.9)   7.0±4.0  
Chromosome          
Good 21 19 (90.5) 0.002* 27.1±3.2 0.000*
Intermediate 26 13 (50.0)   8.3±1.6  
Poor 5 1 (20.0)   2.4±0.4  
mdr1          
Negative 32 26 (81.3) 0.001* 19.9±2.8 0.001*
Positive 20 7 (35.0)   2.0±0.7  
mp1          
Negative 27 21 (77.8) 0.043* 17.0±5.7 0.003*
Positive 25 12 (48.0)   4.0±1.6  
gst π          
Low 21 14 (66.7) 0.762 21.6±3.4 0.296
High 25 15 (60.0)   7.0±3.6  
JNK activity          
Negative 28 20 (71.4) 0.103 16.0±8.8 0.013*
Positive 2 0 (0.0)   2.0±0.0  

* P<0.05.

Abbreviations: See Fig. 1.

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