Elsevier

Gene

Volume 586, Issue 1, 15 July 2016, Pages 123-128
Gene

Research paper
High-throughput screening of Sirtuin family of genes in breast cancer

https://doi.org/10.1016/j.gene.2016.04.023Get rights and content

Highlights

  • ā€¢

    Sirtuin's family of genes shows differential expressions in breast cancer (BC).

  • ā€¢

    SIRT2, SIRT3, and SIRT5 genes were shown to be up-regulated in breast cancer.

  • ā€¢

    SIRT2, SIRT3, SIRT5, SIRT6 and SIRT7 were differentially expressed in BC cell lines.

Abstract

Mammalian Sirtuins have been shown to perform distinct cellular functions and deregulated expression of these genes was reported to be involved in the development of various malignancies including breast cancer. An increasing number of evidence indicates that Sirtuins have both tumor promoter and tumor suppressor functions. However, the roles of Sirtuins have not been well-reported in breast cancer. In the present study, quantitative expression levels of Sirtuins (SIRT1ā€“7) in breast cancer patients and breast cancer cell lines (MCF-7 and SKBR3) and control cell line (CRL-4010) were assessed by using a high-throughput real-time PCR method. As a result, Sirtuins were found to be differentially expressed in breast cancer tissues and cancer cell lines. Particularly, expressions of SIRT1 and SIRT4 were found to be significantly down-regulated in breast cancer tissues and SKBR3 breast cancer cells. In contrast, SIRT2, SIRT3, and SIRT5 genes were shown to be up-regulated in our study. Although SIRT6 and SIRT7 were also up-regulated in breast cancer tissues, these expression changes were statistically insignificant. Additionally, SIRT2, SIRT3, SIRT5, SIRT6 and SIRT7 were found to be differentially expressed in breast cancer cell lines. Yet, these changes were not well-correlated with tissue expression levels. In conclusion, Sirtuin family of genes shows differential expressions in breast cancer tissues and cells and SIRT1 and SIRT4 seem to play key tumor suppressor roles in breast cancer development. Herein, we report expression levels of Sirtuin family of genes in both breast cancer tissues and cancer cell lines simultaneously.

Introduction

Breast cancer is a severe malignant disease with poor survival rates and predominantly affects female population (Yumrutas et al., 2015). So far, a tremendous effort has been made to identify novel genetic corner stones with diagnostic, prognostic, and therapeutic potential (Ergun et al., 2015a). However, the precise role of Sirtuin family of genes has not been well reported.

Sirtuins are family of nicotinamide adenine dinucleotide (NADĀ +)-dependent protein modifying enzymes with certain enzymatic activities including lysine deacetylation, ADP-ribosylation, and/or deacylation (Roth and Chen, 2014). Among four classes of histone deacetylases (HDACs), Sirtuins are classified as class III HDACs with their distinct NADĀ +-dependency (Yuan et al., 2013). Mammalian Sirtuins comprise seven members known as SIRT1ā€“7 and each have distinct enzymatic activities (Roth and Chen, 2014, Yuan et al., 2013). The well-known member of this family is SIRT1 which has been reported to have longevity-promoting effects similar to its homolog yeast silent information regulator 2 (Sir2) (Roth and Chen, 2014, Yuan et al., 2013). More recently, members of this protein family have been implicated to have diverse roles in the cancers including breast carcinogenesis (Yuan et al., 2013). SIRT1 was shown to have dual functions in tumor development (Yuan et al., 2013). Likewise, a mass of evidence also suggests that SIRT2 has both tumor suppressor and promoter functions. Particularly, SIRT2 expression was shown to be reduced in gliomas, esophageal adenocarcinomas, gastric adenocarcinomas, and HNSCC (Hiratsuka et al., 2003, Peters et al., 2010). In contrast, silencing SIRT2 was shown to trigger apoptosis of HeLa and C6 glioma cells (He et al., 2012, Li et al., 2011). SIRT3 is known as mitochondrial tumor suppressor and its expression was found to be diminished in human breast and colon carcinoma (Finley et al., 2011), HNSCC (Lai et al., 2013), HCC, and osteosarcoma (Zhang et al., 2012, Zhang and Zhou, 2012). Moreover, the precise role of SIRT4 in cancer formation is still unclear and remains elusive. Yet, there are few reports that showed SIRT4 as a possible tumor suppressor (Jeong et al., 2013, Csibi et al., 2013). In addition, growing evidence indicates SIRT6 as a tumor suppressor and it's reported to be down-regulated in several types of malignancies including pancreas, colorectal, and hepatocellular cancers. On the other hand, some other evidences indicate that it may have tumor promoter functions (Wang et al., 2011, Khongkow et al., 2013). Lastly, expression levels of SIRT7 were shown to be elevated in breast and thyroid cancers (De Nigris et al., 2002). However, the precise role of Sirtuin family of genes in breast carcinogenesis is not undoubtedly understood and remains elusive.

Accordingly, in the present work, the roles of Sirtuin family of genes were assessed by using a high-throughput real-time PCR method. In particular, expression levels of SIRT1ā€“7 were evaluated in both breast cancer tumor samples and cell lines by using Fluidigm BioMarkā„¢ HD System Real-Time PCR.

Section snippets

Study population and collection of tissue samples

A total of 71 breast cancer patients were enrolled for the study. 7 patients were excluded from the study due to the lack of expression data and/or absence of clinical and demographic characteristics. Ethical approval for the present study was taken from the local ethics committee of University of Gaziantep (Ethical approval number: 11.01.2016/3) in compliance with Helsinki Declaration. All study participants were given an informed consent prior to inclusion to study. Fresh tumoral and adjacent

Differential expressions of Sirtuins in breast cancer tissues

To investigate the precise role of Sirtuin family of genes we used a high-throughput Real-Time PCR approach. As a result, expression levels of SIRT2 (pĀ <Ā 0.0001), SIRT3 (pĀ =Ā 0.0078) and SIRT5 (pĀ =Ā 0.002) were found to be significantly elevated in breast cancer tissues as compared to adjacent normal breast tissues. Although SIRT6 and SIRT7 genes were also up-regulated in breast cancer tissues, these changes were shown to be statistically insignificant (pĀ >Ā 0.05). Additionally, expressions of SIRT1 (pĀ <Ā 

Discussion

In this particular study, expression levels of Sirtuin genes were reported in both breast cancer tissues and cell lines. This is the first report that extensively includes both tissue and cell lines expression levels of Sirtuin genes in breast cancer at the same time. Also, expression levels of some of the Sirtuin target genes were also presented (Supplementary Fig. 1).

Studies showed that Sirtuins play important roles in a variety of biological processes including cancer. Among Sirtuins, SIRT1

Conclusions

In conclusion, emerging roles of Sirtuins in cancer formation are expanding rapidly and accumulating results indicate that this gene family has crucial roles in the development of many tumors including breast tumors. In the present work, Sirtuins were reported to be differentially expressed in breast cancer tissues and cell lines. Our study is the first report that shows expression levels of all Sirtuin family of genes in both breast cancer tissues and cancer cell lines. Indeed, Sirtuins seem

Conflict of interest

None.

Acknowledgment

None.

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