| Objective:To investigate the expression of micro RNA 145(mi R-145) and c-myc m RNA in bone marrow of acute myeloid leukemia by real-time fluorescent quantitative PCR, to analyze the correlation between mi R-145 and c-myc m RNA expression in AML patients, and to explore its clinical significance.Methods:1.Research objects: We collected Specimens of 70 cases patients with acute myeloid leukemia, including 46 new diagnosed patients, according to FAB classification, M1 of 6 examples, M2 of 11 cases, 11 M3, 7 M4, 6 M5, and M6 of 5cases. 24 patients with complete remission, M1 of 1 examples, 9 M2, 6 M3, 3 M4, 2 M5,and M6 of 3 cases according to FAB classification.14 cases of non-neoplastic patients were used as controls.2.Design and synthesis of primers: We designed software with reference to the literature design mi R-145 and U6 internal gene, c-myc and GAPDH internal gene sequences of primers by Using Premier 5.0.3.After separation of bone marrow mononuclear cells, and then was extracted total RNA. To detect the m RNA expression levels of mi R-145 and c-myc by real-time quantitative PCR, and to calculate the relative expression values of mi R-145 and c-myc.4.The central tendency was responsed by the median M and variables were assessed by the Mann-Whitney U rank-sum test. The correlation analysis between mi R-145 and WBC count, hematoglobin count, blood platelet cout, percentage of bone marrow blasts were analyzed by the Spearman correlation test. P value≤0.05 was considered statistically significant.Results:1. The relative expression values of mi R-145 m RNA are skew distribution. The values in new diagnosed AML patients was 0.560(0.004~6.037), in remission group was 5.712(1.323~22.846) and in the control group was 14.547(2.019~41.466). TheV relative expression level of mi R-145 in bone marrow of AML was less than that incontrol group and remission group(P<0.001), and the remission group was significantly lower than that in the control group(P=0.012 < 0.05).2. The relative expression of mi R-145 showed no significantly correlation with the percentage of blasts cells in bone marrow, the hematoglobin count, the blood platelet cout and the percentage of peripheral blood Leukocyte count(P>0.05).3. The relative expression of mi R-145 in 9 cases of acute leukemia patients diagnosed newly was significantly lower than after treatment(P < 0.05).4. Mi R-145 in different subtype of AML patients treated first expression is different, the expression level of mi R-145 with M3 type was 0.639(0.013~5.751), in non-M3 type was 0.545(0.004~41.466), but the differences was not statistically significant(P=0.887>0.05).5. The relative expression values of c-myc m RNA in new diagnosed AML patients was 7.668(0.013~36.098), in the remission group was 0.976(0.076~13.212)and in the control group was 0.190(0.005~2.702). The relative expression level of c-myc m RNA of the new diagnosised group was higher than that in control group and remission group(P<0.001), and the remission group was higher than that in the control group, with statistical difference(P=0.015 < 0.05).6. C-myc in different subtype of AML patients treated first expression is different, the expression level of c-myc with M3 type was 7.48(0.06~23.98), in non-M3 type was 7.75( 0.01~36.10), but the differences was not statistically significant(P=0.78>0.05).7. The correlation analysis of mi R-145 and c-myc m RNA expresison:Significantly negative correlation was found beween the expresison of mi R-145 and c-myc m RNA in the initial group(r=-0.564,p<0.001).Conclusion:1. The reduced level of mi R-145 in AML patients indicating that mi R-145 may take Part in the Pathogenesis of AML.2.The expression of c-myc in bone marrow cells of AML patients is significantly elevated which related with the development of acute leukemia.3.There is significant negative correlation between the expresison of mi R-145 and c-myc.4.There are no correlation between the expressions level of mi R-145 and c-myc with the percentage of bone marrow blast cells, the hematoglobin count, the blood platelet cout, the peripheral blood leukocyte count of AML and the immunophenotyping of first treated AML patients. |
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