论文部分内容阅读
FHIT基因位于染色体 3p14 .2 ,实验室的研究证实FHIT基因具有抑癌基因活性。FHIT基因异常表达或表达缺失出现在多种实体肿瘤和造血系统恶性疾病。为了探讨白血病细胞中FHIT基因的异常表达及意义 ,采用巢式RT PCR法对急性髓性白血病 (AML)、急性淋巴细胞白血病 (ALL)、慢性粒细胞白血病 (CML)以及骨髓增生异常综合症 (MDS)、慢性淋巴细胞白血病 (CLL)、骨髓瘤 (MM )等不同类型白血病患者FHIT基因转录本进行检测 ,并对FHIT基因转录本进行序列测定。 98份血液系统恶性疾病患者的骨髓或外周血标本中AML 38例 ,ALL16例 ,CML 34例 ,其它恶性血液病 10例。全部患者经骨髓细胞形态检查 ,诊断符合FAB诊断标准。肝素抗凝骨髓或外周血 2 - 3ml,分离单个核细胞 ,取 5× 10 7个细胞 ,用RTIZOL 试剂一步法提取细胞总RNA ,行PCR扩增 ,琼脂糖凝胶电泳分析PCR产物。回收目的片段 ,克隆到PGEM T载体 ,进行序列测定。结果表明 :在 5 5 / 98(5 6 % )的被检测病例有FHIT基因的异常表达 ,在AML、ALL和CML异常表达的发生率分别为 2 2 / 38(5 8% ) ,9/ 16(5 6 % )和 19/ 34(5 6 % )。正常骨髓或外周血标本未见有FHIT基因的异常表达。 4 3/ 98(44 % )的患者表达正常转录本 (Ⅰ型 ) ,4 0 / 98(41% )的患者同时表达正常转录本和异常
FHIT gene is located on chromosome 3p14.2, laboratory studies have confirmed that FHIT gene has tumor suppressor gene activity. FHIT gene abnormal expression or lack of expression appears in a variety of solid tumors and hematopoietic malignancies. In order to investigate the abnormal expression and significance of FHIT gene in leukemia cells, a nested reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of FHIT gene in leukemia cells. Methods Acute neutrophil leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML) and myelodysplastic syndrome MDS, CLL and MM were used to detect the FHIT gene transcripts. The FHIT gene transcripts were sequenced. There were 38 cases of AML, 16 cases of ALL, 34 cases of CML and 10 cases of other hematologic malignancies in bone marrow or peripheral blood of 98 patients with hematologic malignancies. All patients by bone marrow cell morphology examination, diagnosis consistent with FAB diagnostic criteria. Peripheral blood of 2 to 3 ml of heparin is used for anticoagulation. Mononuclear cells are isolated and 5 × 10 7 cells are taken. Total RNA is extracted by one-step use of RTIZOL reagent. The PCR products are amplified by PCR and analyzed by agarose gel electrophoresis. The target fragment was recovered and cloned into PGEM T vector for sequencing. The results showed that the abnormal expression of FHIT gene was detected in 55/98 (56%) cases, and the rates of abnormal expression of AML, ALL and CML were 22/38 (58%), 9/16 (56%) and 19/34 (56%). Normal bone marrow or peripheral blood specimens showed no abnormal expression of FHIT gene. Forty-three of 98 (44%) patients expressed normal transcripts (type I) and 40/98 (41%) of patients expressed both normal transcripts and abnormalities