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目的研究抑癌基因、端粒酶及DNA修复基因在白血病细胞中的表达,探讨其临床意义。方法收集儿童急性白血病骨髓活检标本45例(包括初诊组25例和缓解组20例),以及对照组非白血病患儿骨髓标本20例。(1)以骨髓组织石蜡切片作原位杂交PTENmRNA检测,并用免疫组化EliVision法检测端粒酶催化亚单位(hTERT)和DNA依赖蛋白激酶催化亚单位(DNA-PKcs)的表达。比较3组差异。(2)初诊组中20例治疗4周,观察疗效与3项指标表达的关系。结果(1)白血病初诊组骨髓腔充满白血病细胞。PTENmRNA失表达40．0％,hTERT阳性表达84．0％,DNA-PKcs失表达28．0％;3项表达水平均与缓解组和对照组差别显著(P＜0．05)。(2)化学药物治疗4周,未完全缓解者5例;PTENmRNA失表达者占4例,CR率低于阳性表达者(P＜0．05);hTERT和DNA-PKcs阳性表达者各占4例,但无统计学显著差异(P＞0．05)。结论(1)骨髓活检有助于白血病诊断和疗效判定。(2)儿童急性白血病显示有PTENmRNA、hTERT及DNA-PKcs表达异常,表明抑癌基因、端粒酶和DNA修复基因在白血病发生中起到重要作用,可作为基因诊断。(3)白血病初诊组与缓解组显著差别,可辅助疗效判定。(4)治疗效果与多因素相关,PTENmRNA失表达可影响疗效,显示有预测预后价值。
Objective To study the expression of tumor suppressor gene, telomerase and DNA repair gene in leukemia cells and to explore its clinical significance. Methods 45 cases of acute leukemia bone marrow biopsy specimens were collected (including 25 cases of newly diagnosed group and 20 cases of remission group), and 20 cases of bone marrow samples of non-leukemia children in control group. (1) Paraffin sections of bone marrow tissue were used for PTENmRNA in situ hybridization, and the expression of telomerase catalytic subunit (hTERT) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) was detected by immunohistochemistry EliVision. Compare the three groups of differences. (2) In the newly diagnosed group, 20 cases were treated for 4 weeks, and the relationship between the curative effect and the expression of 3 indicators was observed. Results (1) The leukemia newly diagnosed group was filled with leukemia cells in marrow cavity. The loss of PTEN mRNA was 40.0%, the expression of hTERT was 84.0%, and the loss of DNA-PKcs was 28.0%. The expression of PTEN mRNA was significantly different from that of remission group and control group (P <0.05). (2) Chemotherapy 4 weeks, 5 cases of incomplete remission; PTENmRNA loss of expression accounted for 4 cases, CR rate was lower than the positive expression (P <0.05); hTERT and DNA-PKcs positive expression of each 4 Cases, but no statistically significant difference (P> 0.05). Conclusion (1) Bone marrow biopsy can be helpful for diagnosis and treatment of leukemia. (2) The expression of PTENmRNA, hTERT and DNA-PKcs in children with acute leukemia is abnormal, indicating that tumor suppressor gene, telomerase and DNA repair gene play an important role in the genesis of leukemia and can be used as gene diagnosis. (3) Leukemia newly diagnosed group and remission group were significantly different, can be adjuvant efficacy determination. (4) The curative effect is related to many factors. The loss of PTEN mRNA expression can affect the curative effect, indicating the prognostic value.