Dataset Information

Recombinant human decorin suppresses liver HepG2 carcinoma cells by p21 upregulation.

ABSTRACT:

Background

Decorin is a multifunctional molecule of the extracellular matrix and impedes different kinds of tumor cell growth, but the role and molecular mechanism by which decorin inhibits HepG2 cells is not fully understood. Our objective was to construct recombinant human decorin (pcDNA3.1-DCN) and to explore the mechanism by which it inhibits HepG2 cells.

Methods

This experiment was divided into three groups, ie, a control group, an empty vector group, and a pcDNA3.1-DCN group. pcDNA3.1-DCN was constructed using recombinant DNA technology, and the vector for pcDNA3.1-DCN and pcDNA3.1 was then transfected into HepG2 cells using Lipofectamine 2000.

Results

Compared with cells in the control group and in the empty vector group, growth of cells in the pcDNA3.1-DCN group was significantly suppressed, the ratios of cells in the G0/G1 phases and proportion of early apoptotic cells were significantly increased, and the level of p21(WAF1/CIP1) (p21) protein was markedly upregulated (P < 0.05). However, there was no significant difference among the three groups in p53 protein expression (P > 0.05).

Conclusion

The pcDNA3.1-DCN vector was successfully constructed and transfected into HepG2 cells, and decorin overexpression suppressed the growth of HepG2 cells by upregulation of p21 via a p53-independent pathway.

SUBMITTER: Zhang Y

PROVIDER: S-EPMC3422087 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Publications

Recombinant human decorin suppresses liver HepG2 carcinoma cells by p21 upregulation.

Zhang Yucheng Y Wang Yali Y Du Zhenwu Z Wang Qian Q Wu Mei M Wang Xiaofeng X Wang Lingling L Cao Linlin L Hamid Abdu Selim AS Zhang Guizhen G

OncoTargets and therapy 20120813

<h4>Background</h4>Decorin is a multifunctional molecule of the extracellular matrix and impedes different kinds of tumor cell growth, but the role and molecular mechanism by which decorin inhibits HepG2 cells is not fully understood. Our objective was to construct recombinant human decorin (pcDNA3.1-DCN) and to explore the mechanism by which it inhibits HepG2 cells.<h4>Methods</h4>This experiment was divided into three groups, ie, a control group, an empty vector group, and a pcDNA3.1-DCN group ...[more]

PMID: 22927763

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Project description:Mercury is widely distributed in the biosphere, and its toxic effects have been associated with human death and several ailments that include cardiovascular diseases, anemia, kidney and liver damage, developmental abnormalities, neurobehavioral disorders, autoimmune diseases, and cancers in experimental animals. At the cellular level, mercury has been shown to interact with sulphydryl groups of proteins and enzymes, to damage DNA, and to modulate cell cycle progression and/or apoptosis. However, the underlying molecular mechanisms of mercury toxicity remain to be elucidated. Our laboratory has demonstrated that mercury exposure induces cytotoxicity and apoptosis, modulates cell cycle, and transcriptionally activates specific stress genes in human liver carcinoma cells. The liver is one of the few organs capable of regeneration from injury. Dormant genes in the liver are therefore capable of reactivation. In this research, we hypothesize that mercury-induced hepatotoxicity is associated with the modulation of specific gene expressions in liver cells that can lead to several disease states involving immune system dysfunctions. In testing this hypothesis, we used an Affymetrix oligonucleotide microarray with probe sets complementary to more than 20,000 genes to determine whether patterns of gene expressions differ between controls and mercury (1-3 microg/mL) treated cells. There was a clear separation in gene expression profiles between controls and mercury-treated cells. Hierarchical cluster analysis identified 2,211 target genes that were affected. One hundred and eighty-eight of these genes were up-regulated, among which forty eight were significant (p = 0.001) with greater than a two-fold change difference in the concentration range (1-3 microg/mL) of mercury-treated cells; twelve genes were moderately over-expressed with an increase of more than one fold (p = 0.004). 2,023 genes were down-regulated with only forty of them reaching statistically significant decline at p = 0.05 according to the Welch's ANOVA/Welch's t-test. Further analyses of affected genes identified genes located on all human chromosomes with higher than normal effects on genes found on chromosomes 1-14, 17-20 (sex-determining region Y)-box18SRY, 21 (splicing factor, arginine/serine-rich 15 and ATP-binding), and X (including BCL6-co-repressor). These genes are categorized as control and regulatory genes for metabolic pathways involving the cell cycle (cyclin-dependent kinases), apoptosis, cytokine expression, Na+/K+ ATPase, stress responses, G-protein signal transduction, transcription factors, DNA repair as well as metal-regulatory transcription factor 1, MTF1 HGNC, chondroitin sulfate proteoglycan 5 (neuroglycan C), ATP-binding cassette, sub-family G (WHITE), cytochrome b-561 family protein, CDC-like kinase 1 (CLK1 HGNC) (protein tyrosine kinase STY), Na+/H+ exchanger regulatory factor (NHERF HGNC), potassium voltage-gated channel subfamily H member 2 (KCNH2), putative MAPK activating protein (PM20, PM21), ras homolog gene family, polymerase (DNA directed), delta regulatory subunit (50 kDa), leptin receptor involved in hematopoietin/interferon-class (D200- domain) cytokine receptor activity and thymidine kinase 2, mitochondrial TK2 HGNC and related genes. Significant alterations in these specific genes provide new directions for deeper mechanistic investigations that would lead to a better understanding of the molecular basis of mercury-induced toxicity and human diseases that may result from disturbances in the immune system.

Dataset Information

Recombinant human decorin suppresses liver HepG2 carcinoma cells by p21 upregulation.

Background

Methods

Results

Conclusion

Publications

Recombinant human decorin suppresses liver HepG2 carcinoma cells by p21 upregulation.

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