叶绿酸抗细胞恶变相关cDNA序列的克隆

Cloning of cDNA Sequences Related to Chlorophyllin Antitransforming Activity against Anti-BPDE

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。

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.