[Effect of CDK7 gene silencing against hepatoblastoma HepG2 cell proliferation].
OBJECTIVE
To screen the antisense oligodeoxynucleotides (ASODN) that inhibit cultured hepatoblastoma HepG2 cell proliferation, and evaluate the antiproliferative potency of modified ASODN.


METHODS
ASODN sequences were selected based on the secondary structure of human CDK7 mRNA predicted with RNAStructure 3.71 software. The binding thermodynamics of CDK7 mRNA to ASODN was analyzed by OligoWalk program. The sequences with the strongest effect against cultured HepG2 cell proliferation in vitro were selected, and the fragments complementary to 1-5 bases upstream or downstream to the complementary region were structurally modified and screened.


RESULTS
The partial phosphorothioate ASODN complementary to 284-303 region of human CDK7 mRNA was the most powerful inhibitor, and the antiproliferative activity reached 40.4+/-12.6%; in the second round of screening, the antiproliferative activity of the full phosphorothioate ASODN complementary to the 287-306 region of the mRNA on HepG2 cells was 68.3+/-2.6%, with IC50 of 51.9+/-8.6 nmol/L.


CONCLUSIONS
Proliferation of HepG2 cells can be significantly inhibited by the screened ASODN, which might be used as a lead compound in the development of specific CDK7 inhibitors.
Zhao
Ai-guo
AG
Institute of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China.
Wu
Shu-guang
SG
D000255:Adenosine Triphosphate / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018844:Cyclin-Dependent Kinases / Q000235:genetics*; D020868:Gene Silencing; D018197:Hepatoblastoma / Q000235:genetics* / Q000473:pathology; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000473:pathology; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics*; D010766:Phosphorylation; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology; D000095043:Cyclin-Dependent Kinase-Activating Kinase
D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D000255:Adenosine Triphosphate; D018844:Cyclin-Dependent Kinases; D000095043:Cyclin-Dependent Kinase-Activating Kinase; C000722416:CDK7 protein, human