目的:建立分析不同PPP2R5C转录本检测方法,分析正常人和血液肿瘤病人中5种PPP2R5C基因转录本的表达特点。方法:根据PPP2R5C基因的结构特点,设计4对引物,利用RT-PCR方法分析9例正常人、7例T细胞-非霍奇金淋巴瘤(T-NHL)、11例B细胞-急性淋巴细胞白血病(B-ALL)和8例急性髓细胞白血病(AML)病人外周血单个核细胞(PBMC)中PPP2R5C基因的表达情况。结果:所设计的4对引物,均可在样本中扩增到预期PCR产物,其中第1对引物可扩增两个不同大小PCR产物(包含12+13+14外显子的大片段及仅含12+14外显子的小片段),而另3对对引物所扩增的产物分别为10+11+12+12 a外显子,Ⅲ+2外显子和Ⅳ+2外显子的基因片段,所有PCR产物均通过核苷酸序列分析证实。结果显示所有样本均表达全部PPP2R5C转录本。结论:初步建立区分不同PPP2R5C转录本的方法,各转录本在正常人和不同血液肿瘤病人中的分布情况基本一致。
Aim:To establish the method for identification of different transcripts of PPP2R5C,and thereby to analyze the expression feature of PPP2R5C transcripts in leukemia patients and healthy individuals. Methods: Four pairs of primers were designed according to the sequences of different PPP2R5C transcripts.PCR were performed to detect the expression of PPP2R5C gene in peripheral blood mononuclear cells from 9 healthy individuals and 26 hematological malignance patients(7 cases with T-cell lymphoma,11 cases with B-ALL and 8 cases with AML).Results: Expected amplicons were amplified by using the four primer pairs,and two PCR products different of size PCR products can be amplified by using the first primer pairs(the larger one included exons 12+13+14 and the small one included exons 12+14).The products with exons 10+11+12+12a、Ⅲ+2 or Ⅳ+2 were amplified by the other three primer pairs.All the PCR products were confirmed by nucleotide sequencing analysis.All transcripts of PPP2R5C were identified in all samples from both healthy individuals and hematological malignance patients.Conclusion:The RT-PCR method was successfully established for identification of different transcripts of PPP2R5C.A similar expression pattern of PPP2R5C transcripts was found in different hematological malignance and healthy individuals.