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Evaluation of the selenotranscriptome expression in two hepatocellular carcinoma cell lines.

Guariniello S, Di Bernardo G, Colonna G, Cammarota M, Castello G, Costantini S - Anal Cell Pathol (Amst) (2015)

Bottom Line: Hence, it needs to identify always new putative markers to improve its diagnosis and prognosis.In the present paper we have carried out a global analysis of the selenotranscriptome expression in HepG2 and Huh7 cells compared to the normal human hepatocytes by reverse transcription-qPCR (RT-qPCR).Our data showed that in both cells there are three downregulated (DIO1, DIO2, and SELO) and ten upregulated (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1) genes.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Biochimica, Biofisica e Patologia Generale, Seconda Università degli Studi di Napoli, 80138 Napoli, Italy.

ABSTRACT
Hepatocellular carcinoma (HCC) is the most common type of liver cancer and is still one of the most fatal cancers. Hence, it needs to identify always new putative markers to improve its diagnosis and prognosis. Since the selenium is able to fight the oxidative damage which is one of the major origins of cell damage as well as cancer, we have recently focused our attention on selenoprotein family and their involvement in HCC. In the present paper we have carried out a global analysis of the selenotranscriptome expression in HepG2 and Huh7 cells compared to the normal human hepatocytes by reverse transcription-qPCR (RT-qPCR). Our data showed that in both cells there are three downregulated (DIO1, DIO2, and SELO) and ten upregulated (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1) genes. Additionally, interactomic studies were carried out to evaluate the ability of these down- and upregulated genes to interact between them as well as to identify putative HUB nodes representing the centers of correlation able to exercise a direct control over the coordinated genes.

No MeSH data available.


Related in: MedlinePlus

Expression of selenoprotein genes analyzed by means of RT-qPCR. The average value for the expression of these genes was obtained from three independent experiments. (a) shows the downregulated genes in HepG2 and Huh7 versus normal hepatocytes (DIO1, DIO2, and SELO) while (b) shows the ten upregulated genes (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1). In each reaction, the expression levels were normalized to the average of the control gene (18S rRNA) and expressed as arbitrary units. The mRNA levels in hepatocytes, HepG2, and Huh7 were evaluated by using the ΔΔCt method. Significant differences in relative gene expression between hepatocytes and HepG2 or Huh7 are marked by (∗p value < 0.05) and (∗∗p value < 0.01).
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fig1: Expression of selenoprotein genes analyzed by means of RT-qPCR. The average value for the expression of these genes was obtained from three independent experiments. (a) shows the downregulated genes in HepG2 and Huh7 versus normal hepatocytes (DIO1, DIO2, and SELO) while (b) shows the ten upregulated genes (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1). In each reaction, the expression levels were normalized to the average of the control gene (18S rRNA) and expressed as arbitrary units. The mRNA levels in hepatocytes, HepG2, and Huh7 were evaluated by using the ΔΔCt method. Significant differences in relative gene expression between hepatocytes and HepG2 or Huh7 are marked by (∗p value < 0.05) and (∗∗p value < 0.01).

Mentions: The gene expression profiles of HepG2, Huh7, and normal hepatocyte cells by means of RT-qPCR have shown that in the two HCC cell lines there were three downregulated genes (DIO1, DIO2, and SELO) (Figure 1(a)) and ten upregulated genes (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1) (Figure 1(b)). In detail, two of the three downregulated genes showed a statistically significant difference between HepG2 and Huh7 versus hepatocytes. On the other hand, in the group of the ten upregulated genes five of them (GPX4, GPX7, SELK, SELM, and SEP15) have shown a statistically significant upregulation in HepG2. All the other twelve selenotranscripts have appeared unchanged (data not shown).


Evaluation of the selenotranscriptome expression in two hepatocellular carcinoma cell lines.

Guariniello S, Di Bernardo G, Colonna G, Cammarota M, Castello G, Costantini S - Anal Cell Pathol (Amst) (2015)

Expression of selenoprotein genes analyzed by means of RT-qPCR. The average value for the expression of these genes was obtained from three independent experiments. (a) shows the downregulated genes in HepG2 and Huh7 versus normal hepatocytes (DIO1, DIO2, and SELO) while (b) shows the ten upregulated genes (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1). In each reaction, the expression levels were normalized to the average of the control gene (18S rRNA) and expressed as arbitrary units. The mRNA levels in hepatocytes, HepG2, and Huh7 were evaluated by using the ΔΔCt method. Significant differences in relative gene expression between hepatocytes and HepG2 or Huh7 are marked by (∗p value < 0.05) and (∗∗p value < 0.01).
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4493270&req=5

fig1: Expression of selenoprotein genes analyzed by means of RT-qPCR. The average value for the expression of these genes was obtained from three independent experiments. (a) shows the downregulated genes in HepG2 and Huh7 versus normal hepatocytes (DIO1, DIO2, and SELO) while (b) shows the ten upregulated genes (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1). In each reaction, the expression levels were normalized to the average of the control gene (18S rRNA) and expressed as arbitrary units. The mRNA levels in hepatocytes, HepG2, and Huh7 were evaluated by using the ΔΔCt method. Significant differences in relative gene expression between hepatocytes and HepG2 or Huh7 are marked by (∗p value < 0.05) and (∗∗p value < 0.01).
Mentions: The gene expression profiles of HepG2, Huh7, and normal hepatocyte cells by means of RT-qPCR have shown that in the two HCC cell lines there were three downregulated genes (DIO1, DIO2, and SELO) (Figure 1(a)) and ten upregulated genes (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1) (Figure 1(b)). In detail, two of the three downregulated genes showed a statistically significant difference between HepG2 and Huh7 versus hepatocytes. On the other hand, in the group of the ten upregulated genes five of them (GPX4, GPX7, SELK, SELM, and SEP15) have shown a statistically significant upregulation in HepG2. All the other twelve selenotranscripts have appeared unchanged (data not shown).

Bottom Line: Hence, it needs to identify always new putative markers to improve its diagnosis and prognosis.In the present paper we have carried out a global analysis of the selenotranscriptome expression in HepG2 and Huh7 cells compared to the normal human hepatocytes by reverse transcription-qPCR (RT-qPCR).Our data showed that in both cells there are three downregulated (DIO1, DIO2, and SELO) and ten upregulated (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1) genes.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Biochimica, Biofisica e Patologia Generale, Seconda Università degli Studi di Napoli, 80138 Napoli, Italy.

ABSTRACT
Hepatocellular carcinoma (HCC) is the most common type of liver cancer and is still one of the most fatal cancers. Hence, it needs to identify always new putative markers to improve its diagnosis and prognosis. Since the selenium is able to fight the oxidative damage which is one of the major origins of cell damage as well as cancer, we have recently focused our attention on selenoprotein family and their involvement in HCC. In the present paper we have carried out a global analysis of the selenotranscriptome expression in HepG2 and Huh7 cells compared to the normal human hepatocytes by reverse transcription-qPCR (RT-qPCR). Our data showed that in both cells there are three downregulated (DIO1, DIO2, and SELO) and ten upregulated (GPX4, GPX7, SELK, SELM, SELN, SELT, SELV, SEP15, SEPW1, and TrxR1) genes. Additionally, interactomic studies were carried out to evaluate the ability of these down- and upregulated genes to interact between them as well as to identify putative HUB nodes representing the centers of correlation able to exercise a direct control over the coordinated genes.

No MeSH data available.


Related in: MedlinePlus