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Eplin-alpha expression in human breast cancer, the impact on cellular migration and clinical outcome.

Jiang WG, Martin TA, Lewis-Russell JM, Douglas-Jones A, Ye L, Mansel RE - Mol. Cancer (2008)

Bottom Line: Grade-2/3 tumours had significantly lower levels of EPLIN-alpha compared with grade-1 (p = 0.047 and p = 0.046 vs grade-1, respectively).Patients with poor prognosis had a significantly lower levels of EPLIN-alpha compared with those with good prognosis (p = 0.0081).It is concluded that expression of EPLIN-alpha in breast cancer is down-regulated in breast cancer cells and tissues, a change linked to the prognosis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Metastasis and Angiogenesis Research Group, Cardiff University School of Medicine, Heath Park, Cardiff CF14 4XN, UK. jiangw@cf.ac.uk

ABSTRACT

Introduction: To investigate the expression of EPLIN-alpha, epithelial protein lost in neoplasm, in human breast cancer tissues/cells and investigate the cellular impact of EPLIN-alpha on breast cancer cells.

Experimental design: EPLIN-alpha was determined in tumour (n = 120) and normal mammary tissues (n = 32), and cancer cell lines (n = 16). Cell invasion, in vitro and in vivo growth of cells transfected with EPLIN-alpha were evaluated using in vitro invasion assay, in vitro and in vivo tumour model. Cellular migration was analysed using Electric Cell Impedance Sensing assays.

Results: Low level of EPLIN-alpha was seen in tumour tissues. Grade-2/3 tumours had significantly lower levels of EPLIN-alpha compared with grade-1 (p = 0.047 and p = 0.046 vs grade-1, respectively). Patients with poor prognosis had a significantly lower levels of EPLIN-alpha compared with those with good prognosis (p = 0.0081). Patients who developed recurrence and died of breast cancer had significantly lower levels of EPLIN-alpha compared with those who remained disease free (p = 0.0003 and p = 0.0008, respectively) (median follow-up 10 years). Patients with high levels of EPLIN-alpha transcript had a longer survival than those with low levels. Over-expression of EPLIN-alpha in breast cancer cells by way of transfection rendered cells less invasive, less motile and growing at a slower pace in vitro and in vivo. An ERK inhibitor was shown to be able to abolish the effect of EPLIN expression.

Conclusion: It is concluded that expression of EPLIN-alpha in breast cancer is down-regulated in breast cancer cells and tissues, a change linked to the prognosis. EPLIN-alpha acts as a potential tumour suppressor by inhibition of growth and migration of cancer cells.

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EPLIN-α expression in breast cancer tissues and cell lines.a: Immunohistochemical staining of EPLIN-α in normal mammary tissues (left panel) and tumour tissues (right panel) from different patients. The tumour tissues were from patients with different tumour grade. D was a grade-1 tumour (Patient ID 123), E a grade-2 tumour (ID 75a) and F grade-3 tumour (ID 113). Magnification was ×200 for the micrographs. b1: Detection of the EPLIN-α transcript using RT-PCR in a panel of human breast, endothelial and stromal fibroblast cell lines. b2: Detection of the EPLIN-α transcript in a panel of matched normal and tumour tissues. Actin was used as the housekeeping control. c: quantitative analysis of the EPLIN-α transcript in normal and tumour tissues. d: Levels of expression of EPLIN-α transcripts in breast tumour tissues in connection with TNM status (left; * p = 0.0029 vs TNM1 tumours). e: Levels of expression of EPLIN-α in breast tumour tissues in connection with grade (left, * p < 0.05 vs grade 1 tumours) and F: with nodal status (right). Inserts in d, e and f are values of EPLIN-α transcript that have been normalised by CK19 (shown as EPLIN:CK19 ratio).
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Figure 1: EPLIN-α expression in breast cancer tissues and cell lines.a: Immunohistochemical staining of EPLIN-α in normal mammary tissues (left panel) and tumour tissues (right panel) from different patients. The tumour tissues were from patients with different tumour grade. D was a grade-1 tumour (Patient ID 123), E a grade-2 tumour (ID 75a) and F grade-3 tumour (ID 113). Magnification was ×200 for the micrographs. b1: Detection of the EPLIN-α transcript using RT-PCR in a panel of human breast, endothelial and stromal fibroblast cell lines. b2: Detection of the EPLIN-α transcript in a panel of matched normal and tumour tissues. Actin was used as the housekeeping control. c: quantitative analysis of the EPLIN-α transcript in normal and tumour tissues. d: Levels of expression of EPLIN-α transcripts in breast tumour tissues in connection with TNM status (left; * p = 0.0029 vs TNM1 tumours). e: Levels of expression of EPLIN-α in breast tumour tissues in connection with grade (left, * p < 0.05 vs grade 1 tumours) and F: with nodal status (right). Inserts in d, e and f are values of EPLIN-α transcript that have been normalised by CK19 (shown as EPLIN:CK19 ratio).

Mentions: We first analysed the expression pattern of EPLIN-α in paired breast tissues and breast cancer cell lines. EPLIN-α protein was found in the cytoplasmic region of normal mammary epithelial cells. Stromal cells had very little staining (figure 1a). This would indicate, to some degree, that EPLIN-α is primarily located epithelial cells. However, in tumour tissues, EPLIN-α staining in cancer cells was substantially weaker than in normal epithelial cells (figure 1a). The pattern is largely supported by the analysis of the EPLIN-α transcript using conventional PCR (figure 1b2). Tumour tissues did indeed show a weak signal compared with normal tissues. Most breast cancer cell lines showed a weak presence of EPLIN-α transcript. BT474 and MDA MB-468 showed a stronger signal of the transcript (figure 1b1). Interestingly, two of the fibroblast cell lines also showed a weak signal for EPLIN-α. Interestingly, a human endothelial cells, HUVEC and HECV, had little EPLIN-α transcript.


Eplin-alpha expression in human breast cancer, the impact on cellular migration and clinical outcome.

Jiang WG, Martin TA, Lewis-Russell JM, Douglas-Jones A, Ye L, Mansel RE - Mol. Cancer (2008)

EPLIN-α expression in breast cancer tissues and cell lines.a: Immunohistochemical staining of EPLIN-α in normal mammary tissues (left panel) and tumour tissues (right panel) from different patients. The tumour tissues were from patients with different tumour grade. D was a grade-1 tumour (Patient ID 123), E a grade-2 tumour (ID 75a) and F grade-3 tumour (ID 113). Magnification was ×200 for the micrographs. b1: Detection of the EPLIN-α transcript using RT-PCR in a panel of human breast, endothelial and stromal fibroblast cell lines. b2: Detection of the EPLIN-α transcript in a panel of matched normal and tumour tissues. Actin was used as the housekeeping control. c: quantitative analysis of the EPLIN-α transcript in normal and tumour tissues. d: Levels of expression of EPLIN-α transcripts in breast tumour tissues in connection with TNM status (left; * p = 0.0029 vs TNM1 tumours). e: Levels of expression of EPLIN-α in breast tumour tissues in connection with grade (left, * p < 0.05 vs grade 1 tumours) and F: with nodal status (right). Inserts in d, e and f are values of EPLIN-α transcript that have been normalised by CK19 (shown as EPLIN:CK19 ratio).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2553413&req=5

Figure 1: EPLIN-α expression in breast cancer tissues and cell lines.a: Immunohistochemical staining of EPLIN-α in normal mammary tissues (left panel) and tumour tissues (right panel) from different patients. The tumour tissues were from patients with different tumour grade. D was a grade-1 tumour (Patient ID 123), E a grade-2 tumour (ID 75a) and F grade-3 tumour (ID 113). Magnification was ×200 for the micrographs. b1: Detection of the EPLIN-α transcript using RT-PCR in a panel of human breast, endothelial and stromal fibroblast cell lines. b2: Detection of the EPLIN-α transcript in a panel of matched normal and tumour tissues. Actin was used as the housekeeping control. c: quantitative analysis of the EPLIN-α transcript in normal and tumour tissues. d: Levels of expression of EPLIN-α transcripts in breast tumour tissues in connection with TNM status (left; * p = 0.0029 vs TNM1 tumours). e: Levels of expression of EPLIN-α in breast tumour tissues in connection with grade (left, * p < 0.05 vs grade 1 tumours) and F: with nodal status (right). Inserts in d, e and f are values of EPLIN-α transcript that have been normalised by CK19 (shown as EPLIN:CK19 ratio).
Mentions: We first analysed the expression pattern of EPLIN-α in paired breast tissues and breast cancer cell lines. EPLIN-α protein was found in the cytoplasmic region of normal mammary epithelial cells. Stromal cells had very little staining (figure 1a). This would indicate, to some degree, that EPLIN-α is primarily located epithelial cells. However, in tumour tissues, EPLIN-α staining in cancer cells was substantially weaker than in normal epithelial cells (figure 1a). The pattern is largely supported by the analysis of the EPLIN-α transcript using conventional PCR (figure 1b2). Tumour tissues did indeed show a weak signal compared with normal tissues. Most breast cancer cell lines showed a weak presence of EPLIN-α transcript. BT474 and MDA MB-468 showed a stronger signal of the transcript (figure 1b1). Interestingly, two of the fibroblast cell lines also showed a weak signal for EPLIN-α. Interestingly, a human endothelial cells, HUVEC and HECV, had little EPLIN-α transcript.

Bottom Line: Grade-2/3 tumours had significantly lower levels of EPLIN-alpha compared with grade-1 (p = 0.047 and p = 0.046 vs grade-1, respectively).Patients with poor prognosis had a significantly lower levels of EPLIN-alpha compared with those with good prognosis (p = 0.0081).It is concluded that expression of EPLIN-alpha in breast cancer is down-regulated in breast cancer cells and tissues, a change linked to the prognosis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Metastasis and Angiogenesis Research Group, Cardiff University School of Medicine, Heath Park, Cardiff CF14 4XN, UK. jiangw@cf.ac.uk

ABSTRACT

Introduction: To investigate the expression of EPLIN-alpha, epithelial protein lost in neoplasm, in human breast cancer tissues/cells and investigate the cellular impact of EPLIN-alpha on breast cancer cells.

Experimental design: EPLIN-alpha was determined in tumour (n = 120) and normal mammary tissues (n = 32), and cancer cell lines (n = 16). Cell invasion, in vitro and in vivo growth of cells transfected with EPLIN-alpha were evaluated using in vitro invasion assay, in vitro and in vivo tumour model. Cellular migration was analysed using Electric Cell Impedance Sensing assays.

Results: Low level of EPLIN-alpha was seen in tumour tissues. Grade-2/3 tumours had significantly lower levels of EPLIN-alpha compared with grade-1 (p = 0.047 and p = 0.046 vs grade-1, respectively). Patients with poor prognosis had a significantly lower levels of EPLIN-alpha compared with those with good prognosis (p = 0.0081). Patients who developed recurrence and died of breast cancer had significantly lower levels of EPLIN-alpha compared with those who remained disease free (p = 0.0003 and p = 0.0008, respectively) (median follow-up 10 years). Patients with high levels of EPLIN-alpha transcript had a longer survival than those with low levels. Over-expression of EPLIN-alpha in breast cancer cells by way of transfection rendered cells less invasive, less motile and growing at a slower pace in vitro and in vivo. An ERK inhibitor was shown to be able to abolish the effect of EPLIN expression.

Conclusion: It is concluded that expression of EPLIN-alpha in breast cancer is down-regulated in breast cancer cells and tissues, a change linked to the prognosis. EPLIN-alpha acts as a potential tumour suppressor by inhibition of growth and migration of cancer cells.

Show MeSH
Related in: MedlinePlus