摘要
目的 建立BMI- 1基因(theBcell specificsmoloneymurineLeukemiaVirusintegrationsite 1)表达的实时荧光定量PCR,利用该方法在基因转录水平上研究BMI- 1在间质干细胞 (MSCs)及肿瘤细胞中的表达水平。方法 通过RT- PCR,T- A克隆,real timePCR等方法精确测定并比较BMI 1基因在胚胎MSCs、成人MSCs、A549(人肺腺癌细胞株)、SW 480(人结肠腺癌细胞株)、U937(人髓系白血病细胞株)、健康人外周血淋巴细胞以及白血病细胞中表达水平。结果 测定绘制BMI 1基因的标准曲线(相关系数r=0. 998),胚胎MSCs中BMI- 1的表达水平高于成人MSCs,慢性粒细胞白血病(CML)和慢性淋巴细胞白血病(CLL)中BMI- 1的表达量分别高于急性粒细胞白血病和急性淋巴细胞白血病,U937和A549中的表达量高于SW 480,健康人外周血淋巴细胞中BMI 1的表达明显低于慢性髓系白血病(P<0. 01)。结论 定量测定BMI -1的real- timePCR方法较稳定、准确。BMI- 1基因可以作为某些肿瘤特别是白血病治疗的靶点,其表达水平可作为判断肿瘤恶性程度的参考指标。
Objective To establish a method to detect the expression of BMI-1 gene by real-time fluorescent quantitative PCR,and to investigate the expression of BMI-1 gene at a transcriptional level in mesenchymal stem cells and various tumor cells with the established method.Methods The exact copy number of BMI-1 gene and expression level in embryonic and adult MSCs,different tumor cell lines,e.g.,U937,A549 and SW480,bone marrow cells extracted from leukemia patients were determined by RT-PCR,T-A clone and real-time PCR,and the results were compared each other.Results The standard quantitative curve for BMI-1 gene was generated (correlation coefficient r=0.998).The expression level of BMI-1 in embryonic MSCs was higher than that in adult MSCs,in chronic leukemia cells was higher than that in acute leukemia cells,in U937 and A549 cells was higher than that in SW480 cells,while in PBL(peripheral blood lymphocyte) of healthy subjects was significantly lower than that in chronic myeloblastic leukemia cells.Conclusions A stable,accurate real-time PCR experimental method was established for detection of the lever of BMI-1 gene expression in laboratory.The transcriptional expression difference of BMI-1 gene between MSCs and various tumor cells was tentatively investigated and analyzed.It is likely that the difference may relate to the growth, development and histological type of bone marrow MSCs and other tumors. BMI-1 gene can be regarded as a potential target in therapeutic intervention for tumor,especially for leukemia,and its expression lever should be a useful marker in estimation for malignance of tumor and evaluation for the effect of tumor therapy.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2005年第2期114-116,共3页
Chinese Journal of Clinical Laboratory Science
基金
国家自然科学基金(30471938)
江苏省自然科学基金(BK20002006)
江苏大学高级人才启动基金(045DG028)
