[Purification of a new phospholipase A2 homologue from Agkistrodon blomhoffii siniticus and its effects on gene expression profile of Hep3B cells].
OBJECTIVE
To isolate and purify a new phospholipase A2 (PLA2) homologue from Agkistrodon blomhoffii siniticus and investigate its effects on the gene expression profile of Hep3B cells.


METHODS
The PLA2 homologue was isolated and purified by reverse-phase high-performance liquid chromatography (HPLC) and its purity was determined also by HPLC. The relative molecular mass of the homologue was measured by electrospray ionization mass spectrum. The gene expression profile of Hep3B cells was detected with gene chip after exposure of the cells to 139 microg/ml PLA2 homologue for 12 h.


RESULTS
The purity of the PLA2 homologue was 97.2%, whose relative molecular mass was 13,900. After exposure of Hep3B cells to 139 microg/ml PLA2 homologue for 12 h, 19 genes were down-regulated and 20 up-regulated in the cells. The genes showing altered expressions in response to the exposure were mainly involved in cell cycle control and DNA damage repair, cell apoptosis and senescence, production of signal transduction molecules and transcription factors, cell adhesion, angiogenesis, and tumor invasion and metastasis.


CONCLUSIONS
The PLA2 homologue induces alterations in the expression of a wide variety of genes involved in the growth and metastasis of tumor cells. The results of this study provide clues for further study of the possible mechanism for the action of PLA2 homologue on Hep3B cells.
Ma
An-de
AD
Instrumental Analysis and Research Center, Southern Medical University, Guangzhou 510515, China. mandmy@fimmu.com
Wu
Shao-yu
SY
Zhang
Jia-jie
JJ
Li
Zhi-qin
ZQ
Xu
Wei
W
Wen
Xiao-yun
XY
Yu
Le
L
Wu
Shu-guang
SG
D017836:Agkistrodon; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D004249:DNA Damage / Q000187:drug effects; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D018960:Hyaluronan Receptors / Q000096:biosynthesis / Q000235:genetics; D007527:Isoenzymes; D008113:Liver Neoplasms / Q000235:genetics*; D010741:Phospholipases A / Q000302:isolation & purification / Q000494:pharmacology*; D054467:Phospholipases A2; D051562:Proto-Oncogene Proteins c-bcr / Q000096:biosynthesis / Q000235:genetics; D012910:Snake Venoms / Q000201:enzymology*; D014407:Tumor Cells, Cultured
D018960:Hyaluronan Receptors; D007527:Isoenzymes; D012910:Snake Venoms; C496613:BCR protein, human; D051562:Proto-Oncogene Proteins c-bcr; D010741:Phospholipases A; D054467:Phospholipases A2