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ROCK1 as a novel prognostic marker in vulvar cancer.

Akagi EM, Lavorato-Rocha AM, Maia Bde M, Rodrigues IS, Carvalho KC, Stiepcich MM, Baiocchi G, Sato-Kuwabara Y, Rogatto SR, Soares FA, Rocha RM - BMC Cancer (2014)

Bottom Line: In each pair of samples, ROCK1 levels were higher by qRT-PCR in normal tissue compared with the tumour samples (p = 0.016).ROCK1 is lower expressed in tumour tissue when compared with adjacent normal vulvar epithelia.In an independent sample set of VSCCs, lower expression levels of ROCK1 correlated with worse survival rates and a poor prognosis.

View Article: PubMed Central - PubMed

Affiliation: Molecular Morphology Laboratory, Investigative Pathology, AC Camargo Cancer Center, São Paulo, SP, Brazil. rafael.malagoli@gmail.com.

ABSTRACT

Background: Vulvar carcinoma is an infrequent tumour, accounting for fewer than 3% of all malignant tumours that affect women, but its incidence is rising in the past few decades. In young women, the manifestation of the vulvar carcinoma is often linked to risk factors such as smoking and HPV infection, but most cases develop in women aged over 50 years through poorly understood genetic mechanisms. Rho-associated coiled-coil-containing protein kinase 1 (ROCK1) has been implicated in many cellular processes, but its function in vulvar cancer has never been examined. In this study, we aimed to determine the prognostic value of ROCK1 gene and protein analysis in vulvar squamous cell carcinoma (VSCC).

Methods: ROCK1 expression levels were measured in 16 vulvar tumour samples and adjacent normal tissue by qRT-PCR. Further, 96 VSCC samples were examined by immunohistochemistry (IHC) to confirm the involvement of ROCK1 in the disease. The molecular and pathological results were correlated with the clinical data of the patients. Sixteen fresh VSCC samples were analyzed by array-based comparative genomic hybridization (aCGH).

Results: In each pair of samples, ROCK1 levels were higher by qRT-PCR in normal tissue compared with the tumour samples (p = 0.016). By IHC, 100% of invasive front areas of the tumour and 95.8% of central tumour areas were positive for ROCK1. Greater expression of ROCK1 was associated with the absence of lymph node metastasis (p = 0.022) and a lower depth of invasion (p = 0.002). In addition, higher ROCK1 levels correlated with greater recurrence-free survival (p = 0.001). Loss of ROCK1 was independently linked to worse cancer-specific survival (p = 0.0054) by multivariate analysis. This finding was validated by IHC, which demonstrated enhanced protein expression in normal versus tumour tissue (p < 0.001). By aCGH, 42.9% of samples showed a gain in copy number of the ROCK1 gene.

Conclusions: ROCK1 is lower expressed in tumour tissue when compared with adjacent normal vulvar epithelia. In an independent sample set of VSCCs, lower expression levels of ROCK1 correlated with worse survival rates and a poor prognosis. These findings provide important information for the clinical management of vulvar cancer.

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Related in: MedlinePlus

Representative image of aCGH analysis of chromosome 18 with emphasis onROCK1.(A) The copy number gain (chr18:18,539,853-19,429,001) is in blue; exons of ROCK1 are illustrated at the bottom of the diagram. (B) Example highlighting the gains (≥0.3) in blue and high gains (≥0.6) in yellow. Gene regions covered by each probe can be seen as small dots.
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Fig5: Representative image of aCGH analysis of chromosome 18 with emphasis onROCK1.(A) The copy number gain (chr18:18,539,853-19,429,001) is in blue; exons of ROCK1 are illustrated at the bottom of the diagram. (B) Example highlighting the gains (≥0.3) in blue and high gains (≥0.6) in yellow. Gene regions covered by each probe can be seen as small dots.

Mentions: By aCGH analysis, 29 regions underwent significant copy number alterations: 9 were associated with copy number loss (8p23.3, 5q11.1-q11.2, 3p11.1-q11.1, 9p23, 21p11.2-p11.1, Xq28, 7q36.3, 19p13.3, and 21p11.2), and 20 had gains (1q22, 20q11.21-q11.23, 1p36.23-p36.22, 11q13.3, 19q13.12, 19q13.32, 7q11.21, 7q11.22, 7q11.23, 7q11.23, 7q22.1, 11q12.2-q12.3, 11q12.3, 11q13.2, 16q22.1, 18q11.1-q11.2, 18q11.2, 7p22.2-p22.1, 12q24.31, and 15q11.1-q11.2). Of the latter, region 18q11.1-q11.2, which harbors ROCK1, had more copies than the reference DNA in 42.9% of samples (Figure 5).Figure 5


ROCK1 as a novel prognostic marker in vulvar cancer.

Akagi EM, Lavorato-Rocha AM, Maia Bde M, Rodrigues IS, Carvalho KC, Stiepcich MM, Baiocchi G, Sato-Kuwabara Y, Rogatto SR, Soares FA, Rocha RM - BMC Cancer (2014)

Representative image of aCGH analysis of chromosome 18 with emphasis onROCK1.(A) The copy number gain (chr18:18,539,853-19,429,001) is in blue; exons of ROCK1 are illustrated at the bottom of the diagram. (B) Example highlighting the gains (≥0.3) in blue and high gains (≥0.6) in yellow. Gene regions covered by each probe can be seen as small dots.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4232714&req=5

Fig5: Representative image of aCGH analysis of chromosome 18 with emphasis onROCK1.(A) The copy number gain (chr18:18,539,853-19,429,001) is in blue; exons of ROCK1 are illustrated at the bottom of the diagram. (B) Example highlighting the gains (≥0.3) in blue and high gains (≥0.6) in yellow. Gene regions covered by each probe can be seen as small dots.
Mentions: By aCGH analysis, 29 regions underwent significant copy number alterations: 9 were associated with copy number loss (8p23.3, 5q11.1-q11.2, 3p11.1-q11.1, 9p23, 21p11.2-p11.1, Xq28, 7q36.3, 19p13.3, and 21p11.2), and 20 had gains (1q22, 20q11.21-q11.23, 1p36.23-p36.22, 11q13.3, 19q13.12, 19q13.32, 7q11.21, 7q11.22, 7q11.23, 7q11.23, 7q22.1, 11q12.2-q12.3, 11q12.3, 11q13.2, 16q22.1, 18q11.1-q11.2, 18q11.2, 7p22.2-p22.1, 12q24.31, and 15q11.1-q11.2). Of the latter, region 18q11.1-q11.2, which harbors ROCK1, had more copies than the reference DNA in 42.9% of samples (Figure 5).Figure 5

Bottom Line: In each pair of samples, ROCK1 levels were higher by qRT-PCR in normal tissue compared with the tumour samples (p = 0.016).ROCK1 is lower expressed in tumour tissue when compared with adjacent normal vulvar epithelia.In an independent sample set of VSCCs, lower expression levels of ROCK1 correlated with worse survival rates and a poor prognosis.

View Article: PubMed Central - PubMed

Affiliation: Molecular Morphology Laboratory, Investigative Pathology, AC Camargo Cancer Center, São Paulo, SP, Brazil. rafael.malagoli@gmail.com.

ABSTRACT

Background: Vulvar carcinoma is an infrequent tumour, accounting for fewer than 3% of all malignant tumours that affect women, but its incidence is rising in the past few decades. In young women, the manifestation of the vulvar carcinoma is often linked to risk factors such as smoking and HPV infection, but most cases develop in women aged over 50 years through poorly understood genetic mechanisms. Rho-associated coiled-coil-containing protein kinase 1 (ROCK1) has been implicated in many cellular processes, but its function in vulvar cancer has never been examined. In this study, we aimed to determine the prognostic value of ROCK1 gene and protein analysis in vulvar squamous cell carcinoma (VSCC).

Methods: ROCK1 expression levels were measured in 16 vulvar tumour samples and adjacent normal tissue by qRT-PCR. Further, 96 VSCC samples were examined by immunohistochemistry (IHC) to confirm the involvement of ROCK1 in the disease. The molecular and pathological results were correlated with the clinical data of the patients. Sixteen fresh VSCC samples were analyzed by array-based comparative genomic hybridization (aCGH).

Results: In each pair of samples, ROCK1 levels were higher by qRT-PCR in normal tissue compared with the tumour samples (p = 0.016). By IHC, 100% of invasive front areas of the tumour and 95.8% of central tumour areas were positive for ROCK1. Greater expression of ROCK1 was associated with the absence of lymph node metastasis (p = 0.022) and a lower depth of invasion (p = 0.002). In addition, higher ROCK1 levels correlated with greater recurrence-free survival (p = 0.001). Loss of ROCK1 was independently linked to worse cancer-specific survival (p = 0.0054) by multivariate analysis. This finding was validated by IHC, which demonstrated enhanced protein expression in normal versus tumour tissue (p < 0.001). By aCGH, 42.9% of samples showed a gain in copy number of the ROCK1 gene.

Conclusions: ROCK1 is lower expressed in tumour tissue when compared with adjacent normal vulvar epithelia. In an independent sample set of VSCCs, lower expression levels of ROCK1 correlated with worse survival rates and a poor prognosis. These findings provide important information for the clinical management of vulvar cancer.

Show MeSH
Related in: MedlinePlus