【摘要】 目的建立检测pokemon基因表达的实时荧光定量PCR的方法,用以研究pokemon在白血病细胞、多种肿瘤细胞株及胃癌组织中的表达水平。方法通过RT-PCR、T-A克隆、real-time PCR等方法精确测定并比较pokemon基因在白血病细胞、人结肠癌细胞株(SW480)、人胃癌细胞株(SGC)、人肺癌细胞株(LTEP)、人髓系白血病细胞株(U937)、人前列腺癌细胞株(PC3)、人宫颈癌细胞株(HELA)、人内皮细胞株(UEVC-304)、健康人外周血单个核细胞(PBMC)、胃癌组织中的表达水平。绘制pokemon基因real-time PCR的标准曲线。结果急性淋巴细胞白血病(ALL)中pokemon的表达显著高于急性粒细胞白血病(AML)和慢性粒细胞白血病(CML)及健康人PBMC(P<0.05);SW480中的表达量明显高于SGC、LTEP、U937、PC3、HELA和UEVC-304(P<0.05);胃癌组织表达量高于胃癌旁组织,阳性率为5/7。结论建立的测定pokemon基因表达的real-timePCR方法稳定、准确,pokemon基因可以作为某些肿瘤的一个新的靶点,其表达水平的检测可作为判断某些肿瘤发生的参考指标。更多还原

【Abstract】 Objective To establish a method of real-time fluorescent quantitative PCR to detect the expression of Pokemon gene,and investigate the expression of Pokemon gene in leukemia cells,various tumer cell lines and gastric cancer cells.Methods The exact copy number of Pokemon gene and expression level in different cell lines,e.g.,SW480,SGC,LTEP,U937,PC3,HELA,UEVC-304,bone marrow cells extracted from leukemia patients and the tissue from gastric cancer patients were detemined by RT-PCR,T-A clone and real-time PCR.Results The standard quantitative curve of Pokemon gene was generated(correlation coefficient r=0.994).The expression of Pokemon gene in acute lymphocytic leukemia was significantly higher than that in acute granulocytic leukemia,chronic granulocytic leukemia and peripheral blood mononuder cells(PBMC)of healthy subjects(P<0.05).The expression of Pokemon gene in SW480 cells was significantly higher than that in SGC cells,LTEP cells,U937 cells,PC3 cells,HELA cells and UEVC-304 cells(P<0.05).The expression of Pokemon gene in the tissue of gastric cancer cells was higher than that of relative normal tissue with positive rate of 5/7.Conclusions A stable,accurate real-time PCR was established for detection of Pokemon gene expression.Pokemon gene can be regarded as a new potential target in therapeutic intervention for tumor,and its expression level shoud be a useful marker in estimation for tumorigenesis.更多还原