[The biological function of peroxiredoxin II on Hep3B cells and its underlying mechanism].
OBJECTIVE
To study the biological function and its possible underlying mechanism of peroxiredoxin II (PrxII) in liver cancer cell line Hep3B.


METHODS
Two pairs of double-stranded small interfering RNA (siRNA) targeted on PrxII gene were transfected into Hep3B cells using LipofectamineTM 2000. After confirming the inhibited effects of these siRNAs through Quant SYBR Green polymerase chain reaction and Western blot, the biological characters of Hep3B cell were analyzed by flow cytometry analysis, MTT and colony formation assays. Furthermore, dichlorodihydrofluorescein diacetate (DCFH-DA) and thiobarbituric acid (TBA) assays, for measuring the products of oxidative reaction, such as the reactive oxygen species (ROS) and malondialdehyde (MDA), were applied to explore whether the antioxidant mechanism was involved in the effects of PrxII functioning on Hep3B cell.


RESULTS
The two pairs of siRNA significantly inhibited PrxII mRNA and protein expression. Compared to the mock and blank control groups, the two PrxII-silent groups showed decreased rates of cell growth and clone formation and increased rates of cell apoptosis. The numbers of the formed colonies were 42.0+/-2.8 and 40.5+/-0.7 respectively in the two PrxII-silent groups, while they were 121.5+/-2.1 and 130.0+/-1.4 in the mock and blank control groups (P less than 0.05). The levels of endogenous ROS and MDA were significantly higher in the two PrxII-silent groups than those in the mock and blank control groups (P less than 0.05).


CONCLUSIONS
PrxII might play an important role in the hepatocarcinogenesis, possibly through an antioxidant function which may provide a favorable microenvironment for cancer cell survival and progression.
Yue
Hai-ying
HY
Department of Experimental Pathology, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China.
Dai
Zhi
Z
Guo
Kun
K
Sun
Rui-xia
RX
Cao
Ji
J
Cui
Jie-feng
JF
Su
Jian-jia
JJ
Yang
Chun
C
Ou
Chao
C
Yue
Hui-fen
HF
Li
Yuan
Y
Liu
Yin-kun
YK
D045744:Cell Line, Tumor; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D018384:Oxidative Stress; D054464:Peroxiredoxins / Q000235:genetics*; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D015398:Signal Transduction; D014162:Transfection
D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D054464:Peroxiredoxins