Walid Sasi

Higher expression levels of SOCS 1,3,4,7 are associated with earlier tumour stage and better clinical outcome in human breast cancer

BackgroundSuppressors of cytokine signaling (SOCS) are important negative feedback regulators of the JAK/STAT signaling pathway, and have been recently investigated for their role in the development of different cancers. In this study, we examined the expression of SOCS1-7 genes in normal and breast cancer tissue and correlated this with several clinico-pathological and prognostic factors.MethodsSOCS1-7 mRNA extraction and reverse transcription were performed on fresh frozen breast cancer tissue samples (n = 127) and normal background breast tissue (n = 31). Transcript levels of expression were determined using real-time PCR and analyzed against TNM stage, tumour grade and clinical outcome over a 10 year follow-up period.ResultsSOCS1,4,5,6 and 7 expression decreased with increased TNM stage (TNM1 vs. TNM3 p = 0.039, TNM1 vs. TNM4 p = 0.016, TNM2 vs. TNM4 p = 0.025, TNM1 vs. TNM3 p = 0.012, and TNM1 vs. TNM3 p = 0.044 respectively). SOCS2 and 3 expression decreased with increased Nottingham Prognostic Index (NPI) (NPI1 vs. NPI3 p = 0.033, and NPI2 vs. NPI3 p = 0.041 respectively). SOCS7 expression decreased with higher tumour grade (Grade 3 vs. Grade 2 p = 0.037). After a median follow up period of 10 years, we found higher levels of SOCS1,2 and 7 expression among those patients who remained disease-free compared to those who developed local recurrence (p = 0.0073, p = 0.021, and p = 0.039 respectively). Similarly, we found higher levels of SOCS 2,4, and 7 expression in those who remained disease-free compared to those who developed distant recurrence (p = 0.022, p = 0.024, and p = 0.033 respectively). Patients who remained disease-free had higher levels of SOCS1 and 2 expression compared to those who died from breast cancer (p = 0.02 and p = 0.033 respectively). The disease free survival (DFS) and overall survival (OS) curves showed that higher levels of SOCS1, 3 and 7 were significant predictors of higher DFS (p = 0.015, p = 0.024 and 0.03 respectively) and OS (p = 0.005, p = 0.013 and p = 0.035 respectively). Higher levels of SOCS 4 were significant in predicting better OS (p = 0.007) but not DFS. Immunohistochemical staining of representative samples showed a correlation between SOCS1, 3, 7 protein staining and the SOCS1, 3, 7 mRNA expression.ConclusionHigher mRNA expression levels of SOCS1, 3, 4 and 7 are significantly associated with earlier tumour stage and better clinical outcome in human breast cancer.

The mRNA expression of SETD2 in human breast cancer: correlation with clinico-pathological parameters

BackgroundSET domain containing protein 2 (SETD2) is a histone methyltransferase that is involved in transcriptional elongation.There is evidence that SETD2 interacts with p53 and selectively regulates its downstream genes. Therefore, it could be implicated in the process of carcinogenesis. Furthermore, this gene is located on the short arm of chromosome 3p and we previously demonstrated that the 3p21.31 region of chromosome 3 was associated with permanent growth arrest of breast cancer cells. This region includes closely related genes namely: MYL3, CCDC12, KIF9, KLHL18 and SETD2. Based on the biological function of these genes, SETD2 is the most likely gene to play a tumour suppressor role and explain our previous findings.Our objective was to determine, using quantitative PCR, whether the mRNA expression levels of SETD2 were consistent with a tumour suppressive function in breast cancer. This is the first study in the literature to examine the direct relationship between SETD2 and breast cancer.MethodsA total of 153 samples were analysed.The levels of transcription of SETD2 were determined using quantitative PCR and normalized against (CK19).Transcript levels within breast cancer specimens were compared to normal background tissues and analyzed against conventional pathological parameters and clinical outcome over a 10 year follow-up period.ResultsThe levels of SETD2 mRNA were significantly lower in malignant samples (p = 0.0345) and decreased with increasing tumour stage.SETD2 expression levels were significantly lower in samples from patients who developed metastasis, local recurrence, or died of breast cancer when compared to those who were disease free for > 10 years (p = 0.041).ConclusionThis study demonstrates a compelling trend for SETD2 transcription levels to be lower in cancerous tissues and in patients who developed progressive disease. These findings are consistent with a possible tumour suppressor function of this gene in breast cancer.

The Role of Suppressors of Cytokine Signalling in Human Neoplasms

Suppressors of cytokine signalling 1–7 (SOCS1–7) and cytokine-inducible SH2-containing protein (CIS) are a group of intracellular proteins that are well known as JAK-STAT and several other signalling pathways negative feedback regulators. More recently several members have been identified as tumour suppressors and dysregulation of their biological roles in controlling cytokine and growth factor signalling may contribute to the development of many solid organ and haematological malignancies. This review explores their biological functions and their possible tumour suppressing role in human neoplasms.

In vitro and in vivo effects of suppressor of cytokine signalling 7 knockdown in breast cancer: the influence on cellular response to hepatocyte growth factor.

Purpose. Suppressor of cytokine signaling 7 (SOCS7) is a member of the SOCS family and is known to interact with phospholipase Cγ-1 (PLCγ-1), a key downstream mediator of the hepatocyte growth factor (HGF)/C-MET axis. Here, we report our observations of the effect of knocking down SOCS7 gene on the behaviour of breast cancer cells both in vitro and in vivo and to elucidate whether this involves HGF/C-MET pathway using the PLCγ-1 blocker U73122. Methods. MCF7 and MDA-MB-231 breast cancer cells were transfected with anti-SOCS7 ribozymal transgene, to create sublines with SOCS7 knockdown. The in vitro growth and migration of the cells were evaluated in basic conditions and with HGF and U73122 treatment using growth assays, scratch-wound, and electrical cell impedance sensing (ECIS) migration assays. MCF7 and MDA-MB-231 in vivo tumour xenograft growth were also studied. Results. Basal in vitro growth and migration of both cellular lines and the in vivo MCF7 xenograft growth were significantly enhanced with SOCS7 knockdown. In vitro HGF treatment has further influenced the growth and migration when SOCS7 gene was knocked-down in both cellular lines (P < 0.05). PLCγ-1 pharmacological inhibition of the HGF/C-MET cascade during their in vitro growth and migration seemed to only occur when SOCS7 gene was knocked down. Conclusions. We report a unique regulatory role for SOCS7 in controlling the malignant behaviour of breast cancer lines MCF7 and MDA-MB-231 in vitro and the MCF7 tumour xenografts in vivo. We also report a regulatory role for SOCS7 during the in vitro HGF-induced growth and migration in these cells as HGF treatment and SOCS7 loss have synergistically enhanced these functions. This SOCS7 knockdown-attributed effect could be due to a precise anti-PLCγ-1 role.

Bilateral Florid Papillomatosis of the Nipple: An Unusual Indicator for Metachronous Breast Cancer Development—A Case Report

Adenoma or florid papillomatosis of the nipple (FPN) is a rare benign disease which has histopathological features similar to those of a mammary papillary carcinoma. Here, we report a rare case of bilateral florid papillomatosis of the nipple and breast cancer, with a literature review.

Recurrent Cystic Lymphangioma of the Breast: Case Report and Literature Review

Cystic lymphangioma is a rare congenital lymphatic malformation. We report a case of a 37-year-old female with recurrent cystic lymphangioma of the breast which has progressively developed during pregnancy and breast feeding.

Abstract P1-04-03: Knocking down Suppressor of Cytokine Signaling 7 in breast cancer: The role in Insulin-like Growth Factor - I/Phospholipase Cγ-1 signaling

BACKGROUND: Suppressor of Cytokine Signaling 7 (SOCS7) is a member of the SOCS family and is known to interact with Phospholipase Cγ-1 (PLCγ-1), one of the Insulin like Growth Factor - I (IGF-I) receptor downstream molecules. In the present study, we sought to examine the effect of knocking down SOCS7 gene on breast cancer cells growth and migration and to elucidate whether this has involved IGF-I — PLCγ-1 signaling using the PLCγ-1 blocker U73122. METHODS: Suitable breast cancer cells (MCF7 and MDA-MB-231) were transfected with anti-SOCS7 ribozymal transgene, to create sublines with SOCS7 knockdown that was verified by RT-PCR. The growth and migration of the cells were evaluated in the presence or absence of IGF-I and PLCγ-1 inhibitor using in vitro growth assay and Electrical Cell Impedance Sensing (ECIS) migration assay. RESULTS: MCF7 ΔSOCS7 and MDA-MB-231 ΔSOCS7 (SOCS7 knockdown) were constructed. Both sublines showed a higher rate of growth compared to control cells with and without IGF-I stimulation. U73122 treatment did not appear to change this growth outcome. Using ECIS migration assay, it was shown that knocking down SOCS7 had a significant positive effect on the migration of MCF7 and MDA-MB-231 cells, and that both IGF-I treatment and SOCS7 knockdown had a synergistic positive influence on their migration (p ΔSOCS7 and MDA-MB-231 ΔSOCS7 migration but not that of the control cells. While SOCS7 has acted to control the IGF-I effect, it appeared to cancel the inhibitory function of U73122, indicating a specific anti - PLCγ-1 role for SOCS7 in IGF-I induced breast cancer cellular migration. CONCLUSION: SOCS7 loss resulted in increased growth and migration of breast cancer cells and this had a synergistic effect on their response to IGF-I. This role could be related to its interaction with PLCγ-1 during the cellular migration but not the growth. It may be possible that SOCS7 acts through different mechanism to control the cellular growth. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-04-03.

Gene Transcripts in the Tumor Suppressor Region of Chromosome 3 (3p21).

Background: There is increasing evidence that the short arm of chromosome 3 has a tumour suppressor region that has been associated with permanent growth arrest of tumour cells. We have identified 9 closely related genes in this region (CCDC12, KLH9, KLH18, MYL3, NBEAL2, PTPN23, SCAP, SETD2, and TESSP2) located which could be possible candidates for tumour suppressor genes (TSG).Our objective was to determine, using quantitative PCR, whether the mRNA expression levels of these genes were consistent with a tumour suppressive function in human breast cancer. Methods: A total of 153 samples were analysed. The levels of transcription of all 9 genes were determined using quantitative PCR and normalized against (CK19).Transcript levels within breast cancer specimens were compared with non-cancerous breast tissue and analyzed against conventional pathological parameters and clinical outcome over a 10 year follow-up period. Results: Quantification of CCDC12, KLH9, KLH18, MYL3, NBEAL2, PTPN23, and SCAP mRNA expression after CK19 normalisation showed no statistically significant difference between malignant and normal breast tissue (p=0.23, 0.46, 0.3, 0.141, 0.8, 0.696, and 0.11 respectively). The mRNA expression of TESSP2 was higher in normal samples compared with malignant lesions (347 vs. 35, p=0.0061) but showed no significant associations with clinical outcome.The levels of SETD2 mRNA were significantly lower in malignant samples (406 vs. 17, p=0.0345) and decreased with increasing tumour stage. SETD2 expression levels were significantly lower in samples from patients who developed metastasis, local recurrence, or died from breast cancer when compared to those who were disease free for > 10 years (p=0.041). Conclusion: These findings are consistent with a possible tumour suppressor function of SETD2 in human breast cancer. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6139.

Suppressors of Cytokine Signaling and Their Role in Human Breast Cancer.

BACKGROUND: Suppressors of cytokine signaling (SOCS) are important negative feedback regulators of the JAK/STAT signaling pathway, and have been recently investigated for their role in the development of different cancers. In this study, we examined the expression of SOCS 1-7 genes in normal and breast cancer tissue and correlated this with several clinico-pathological and prognostic factors. PATIENTS AND METHODS: SOCS 1-7 mRNA extraction and reverse transcription were performed on breast cancer tissue samples (n= 127) and normal background breast tissue (n= 31). Transcript levels of expression were determined using real-time PCR and analyzed against TNM stage, tumour grade and clinical outcome over a 10 year follow-up period. RESULTS: SOCS 1,4,5,6 and 7 expression decreased with increased TNM stage (TNM1 vs. TNM3 p= 0.039, TNM1 vs. TNM4 p= 0.016, TNM1 vs. TNM3 p= 0.012 and TNM1 vs. TNM3 p= 0.044 respectively). SOCS 2 and 3 expression decreased with increased Nottingham Prognostic Index (NPI) (NPI1 vs. NPI3 p=0.033 and NPI2 vs. NPI3 p=0.041 respectively). SOCS-7 expression decreased with higher tumour grade (Grade 3 vs. Grade 2 p=0.037). After a median follow up period of 10 years, we found higher levels of SOCS 1, 2 and 7 expression among those patients who remained disease-free compared to those who developed local recurrence (p= 0.0073, p= 0.021 and p= 0.039 respectively). Similarly, we found higher levels of SOCS 2, 4 and 7 expression in those who remained disease-free compared to those who developed distant recurrence (p= 0.022, p=0.024 and p= 0.033 respectively). Patients who remained disease-free had higher levels of SOCS 1 and 2 expression compared to those who died from breast cancer (p= 0.02 and p=0.033 respectively). The disease free survival (DFS) and overall survival (OS) curves showed that higher levels of SOCS 1, 3 and 7 were significant predictors of higher DFS (p=0.015, p=0.024 and p=0.03 respectively) and OS (p=0.005, p=0.013 and p= 0.035 respectively). Higher levels of SOCS 4 were significant in predicting better OS (p=0.007) but not DFS. CONCLUSION Higher mRNA expression levels of SOCS 1, 3, 4 and 7 are significantly associated with earlier tumour stage and better clinical outcomes in human breast cancer. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3157.