[Construction of subtractive cDNA library of apoptosis-related genes in NB4 cells treated by arsenic trioxide].
OBJECTIVE
Construct the gene library of apoptosis related genes in acute promyelocytic leukemia (APL) cell line NB4 cells treated by arsenic trioxide to clarify the apoptotic mechanism of NB4 cells.


METHODS
APL cell line NB4 cells treated with or without arsenic trioxide for 24 hours. Total RNA was extracted and suppress subtractive hybridization (SSH) was conducted according to the manual. With the cDNA of the apoptosis cells as the tester and that of control cells as the driver, forward and reverse hybridization was performed. Differentially expressed genes were linked with pGEM-Teasy cloning vector and transformed into E. coli DH5alpha. The positive clones were screened by blue and white spot. PCR were used to amplify these genes.


RESULTS
The subtractive cDNA libraries related with apoptosis of NB4 cells were successfully constructed.


CONCLUSIONS
The constructed subtractive libraries are suitable for further study on the functional genes associated with apoptosis ofNB4 cells induced by arsenic trioxide.
Di
Chunhong
C
Medical Laboratory of The First Affiliated Hospital of Medical College, Shihezi University, Shihezi 832008, China. chunhongdi@163.com
Gu
Shaohua
S
Tan
Xiaohua
X
Xian
Lingling
L
Wu
Qihan
Q
Yang
Lei
L
D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000494:pharmacology*; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000032:analysis*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000473:pathology*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D010087:Oxides / Q000494:pharmacology*; D017422:Sequence Analysis, DNA
D001152:Arsenicals; D018076:DNA, Complementary; D010087:Oxides; D000077237:Arsenic Trioxide