【摘要】 目的:克隆叶绿酸抑制人支气管上皮细胞系(16HBE)恶性转化的相关差异表达cDNA序列。方法:以叶绿酸及反式-BPDE共同处理的16HBE细胞为实验细胞,以反式-BPDE单独处理的16HBE细胞为对照细胞,采用cDNA代表性差异分析法,比较2种细胞基因表达的差异,分离叶绿酸抗反式-BP-DE作用的相关差异表达cDNA片段。分离的片段分别与pGEM-T载体连接并转化入JM109细菌中,对质粒DNA进行测序。以BLASTN程序进行GenBank的同源性检索。结果:克隆的5条cDNA序列中,有3条为新基因序列,另2条分别与类S18/S6人核糖体蛋白及alpha-烯醇酶等有同源性。结论:克隆的5条cDNA序列所在的基因可能与叶绿酸抗细胞恶性转化分子机制密切相关。更多还原

【Abstract】 Objective To clone differentially expressed cDNA sequences related to inhibiting malignant transformation of human bronchial epithelial cell line (16HBE) by chlorophyllin. Methods 16HBE cells treated together with chlorophyllin and anti-BPDE were conducted as tester, 16HBE cells treated only with anti-BPDE were conducted as driver,and cDNA Representative Diffenrence Analysis(cDNA RDA) was used to compare the differential gene expression between the two kinds of cells.The cDNA fragments were ligated to pGEM-Tvector and transformed into JM109 bacteria. The plasmid DNA were sequenced and compared with data in GenBank by BLASTN. Results Among the 5 cloned cDNA sequences, three were novel and were registered in dbEST database, two showed sequence homology to alpha-enolase and a new found gene ribosomal protein S18/S6-like.Con-Clusions These 5 cDNA sequences might play important roles in antitransforming effect of chlorophyllin.更多还原