c-FLIP在恶性血液病中的表达及其临床相关性研究

Expression and clinical significance of c-FLIP in hematologic malignancies

目的探讨c-FLIP基因在血液肿瘤中的表达以及临床意义。方法以半定量逆转录聚合酶链反应(RT-PCR)检测22例血液肿瘤骨髓中的c-FLIP m RNA表达,Western-Blot蛋白质印迹(WB)方法检测其c-FLIP蛋白的表达。病例包括急性白血病(AL)12例(其中初治11例及反复复发1例)、完全缓解(CR)后AL 2例、慢性粒细胞性白血病(CML)2例、慢性淋巴细胞性白血病(CLL)2例、多发性骨髓瘤(MM)2例和骨髓增生异常综合征-难治性贫血伴原始细胞增多2型(MDS-RAEB-2)2例。结果初治和复发AL,初治CML、CLL、MDS-RAEB-2、MM患者c-FLIP m RNA和蛋白均表达异常增高。初治AL c-FLIP m RNA及43 k D蛋白的表达显著高于慢性白血病(CL)(P<0.05)。MDS-RAEB-2、MM与ALc-FLIP m RNA的表达差异均无统计学差异(P>0.05),但MDS 55 KD、43KD蛋白表达明显低于AL(P<0.05),而MM无明显差异。对照组m RNA及其蛋白均阴性表达,CR后AL m RNA阴性表达,但其蛋白均有表达,低表达于AL,尤其43 k D明显低于AL(P<0.05)。c-FLIP表达在初治AL患者与其年龄、性别、初诊白细胞数、LDH以及核型、免疫表型无关。AL患者c-FLIP m RNA及其蛋白表达与初治疗效无明显相关性(P>0.05)。结论血液肿瘤患者c-FLIP m RNA及其蛋白表达异常增高,能够反映出恶性血液病患者骨髓造血细胞的凋亡抑制情况,与恶性血液病的类型、疾病的状态以及病程中的临床疗效、预后等都密切相关。

Objective To explore c-FLIP（cellular FLICE inhibitory protein） expression and its clinical significance in hematologic malignancies. Methods Expression level of c-FLIP m RNA and protein in bone marrow mononuclear cells was determined by Semi-Reverse Transcription-Polymerase Chain Reaction（Semi-RT-PCR） and Western blot in 22 patients of hematologic malignancies, which included 11 cases with newly diagnosed acute leukemia（AL）, 1 case of relapsed AL, 2 cases of AL after complete remission（CR）, 2 cases of chronic myelocytic leukemia（CML）, 2 cases of chronic lymphocytic leukemia（CLL）, 2 cases of multiple myeloma（MM） and 2 cases of mycelodysplastic syndrome-refractory anemia with excess blasts-2（MDS-RAEB-2）. Results Expression level of c-FLIP m RNA and protein were higher in patients with newly diagnosed or relapsed AL and newly diagnosed CML, CLL,MDS-RAEB-2, MM. Levels of c-FLIP m RNA and protein of 43 k D were higher in AL than in newly diagnosed chronic leukemia（CL）, P〈0.05. There were no significant differences in m RNA between MDS-RAEB-2, MM and AL（P〉0.05）. The levels of c-FLIP protein of 55 k D and 43 k D were lower in MDS than those in newly diagnosed AL（P〈0.05）. The expression of normal subjects in m RNA and protein were negative. The expression of AL after CR was negative in m RNA, but positive in protein, and lower than AL, especially in protein of 43 k D（P〈0.05）. The expression level of c-FLIP was not associated with age, sex, WBC, LDH, cytogenetic characteristics, immunophenotype（P〉0.05）. There were no significant differences in that of curative effect（P〉0.05）. Conclusion c-FLIP was highly expressed in hematologic malignancies. The level of expression reflected apoptosis of hematologic malignancies, and was closely related with the type and situation of hematologic malig-nancies, chemotherapeutic effects and prognosis.