Construction of subtractive cDNA Library of apoptosis-related genes in NB4 cells treated by arsenic trioxide

Chunhong Di, Shaohua Gu, Xiaohua Tan, Lingling Xian, Qihan Wu, Lei Yang

Research output: Contribution to journalArticlepeer-review

Abstract

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. Method: 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 DH5a. The positive clones were screened by blue and white spot. PCR were used to amplify these genes. Result: The subtractive cDNA libraries related with apoptosis of NB4 cells were successfully constructed. Conclusion: The constructed subtractive libraries are suitable for further study on the functional genes associated with apoptosis of NB4 cells induced by arsenic trioxide.

Original languageEnglish (US)
Pages (from-to)454-457
Number of pages4
JournalZhongguo Zhongyao Zazhi
Volume34
Issue number4
StatePublished - Feb 2009
Externally publishedYes

Keywords

  • Apoptosis
  • Arsenic trioxide
  • Leukemia
  • NB4 cell line
  • Suppress subtractive hybridization

ASJC Scopus subject areas

  • General Pharmacology, Toxicology and Pharmaceutics
  • Pharmacology (medical)
  • Complementary and alternative medicine

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