Abstract
Background
Our previous studies demonstrated that p53-induced gene 11 (PIG11) was involved in arsenic trioxide (As2O3)-induced apoptosis in human gastric cancer MGC-803 cells. Here, we studied further PIG11 expression in human hepatocellular carcinoma (HCC) tissues and cell lines and compared the sensitivity to As2O3-induced cell apoptosis in HepG2 and L-02 cells.
Methods
PIG11 expression in human normal liver tissues, HCC tissues, and cell lines was determined by immunohistochemistry and immunocytochemistry methods, using an anti-human PIG11 antibody. Cell viability was estimated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diplenyltetrazolium bromide (MTT) assay. Cell apoptosis was determined by flow cytometry. Reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blotting were performed to analyze PIG11 mRNA and protein expression in cells. Protein intensity was calculated by comparison with the intensity of β-actin, using densitometry. PIG11 was knocked down using small interfering RNA (siRNA).
Results
We found that PIG11 expression was significantly downregulated in HCC tissue and the cell lines (Bel-7402, SMMC-7721, HepG2 cells). Further, HepG2 cells were more sensitive to As2O3-induced apoptosis than L-02 cells. The expression of PIG11 mRNA and protein was upregulated to a greater extent in HepG2 than in L-02 cells. In the presence of actinomycin D or cycloheximide, the amount of PIG11 protein expression did not increase. Likewise, the inhibition of PIG11 by siRNA decreased As2O3-induced PIG11 protein expression by more than 85% and partially prevented As2O3-induced apoptosis in both HepG2 and L-02 cells.
Conclusion
The above results demonstrated that the PIG11 gene may be involved in As2O3-induced apoptosis in HepG2 cells and suggested that the adaptive response of PIG11 expression is one of the important factors in enhancing cell sensitivity to As2O3-induced apoptosis.
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Acknowledgments
This work was supported by grants from the National Science Foundation of China (30370386, 30570461) and the Hunan Provincial Natural Science Foundation of China (06JJ2048).
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Liu, XM., Xiong, XF., Song, Y. et al. Possible roles of a tumor suppressor gene PIG11 in hepatocarcinogenesis and As2O3-induced apoptosis in liver cancer cells. J Gastroenterol 44, 460–469 (2009). https://doi.org/10.1007/s00535-009-0030-1
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DOI: https://doi.org/10.1007/s00535-009-0030-1