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CCL25-CCR9 interaction modulates ovarian cancer cell migration, metalloproteinase expression, and invasion.

Johnson EL, Singh R, Singh S, Johnson-Holiday CM, Grizzle WE, Partridge EE, Lillard JW - World J Surg Oncol (2010)

Bottom Line: OvCa tissue microarrays (TMA) was used to confirm CCR9 expression in clinical samples.Our results show significantly (p<0.001) higher expression of CCR9 by mucinous adenocarcinoma, papillary serous carcinoma, and endometriod ovarian carcinoma cases, than compared to non-neoplastic ovarian tissue.Expression of collagenases (MMP-1, -8, and -13), gelatinases (MMP-2 and -9), and stromelysins (MMP-3, -10, and -11) by OvCa cells were modulated by CCL25 in a CCR9-dependent fashion.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Biochemistry, & Immunology, Morehouse School of Medicine, 720 Westview Drive SW, Atlanta, GA 30310-1495, USA.

ABSTRACT

Background: Ovarian carcinoma (OvCa) is the most lethal gynecological malignancy among women and its poor prognosis is mainly due to metastasis. Chemokine receptor CCR9 is primarily expressed by a small subset of immune cells and its only natural ligand, CCL25, is largely expressed in the thymus, which involutes with age. Other than the thymus, CCL25 is expressed by the small bowel. Interactions between CCL25 and CCR9 have been implicated in leukocyte trafficking to the small bowel, a frequent metastatic site for OvCa cells. The current study shows OvCa tissue and cells significantly express CCR9, which interacts with CCL25 to support carcinoma cell migration and invasion.

Methods: RT-PCR and flow cytometry techniques were used to quantify the expression CCR9 by OvCa cells. OvCa tissue microarrays (TMA) was used to confirm CCR9 expression in clinical samples. The Aperio ScanScope scanning system was used to quantify immunohistochemical staining. Cell invasion and migration assays were performed using cell migration and matrigel invasion chambers. Matrix metalloproteinase (MMP) mRNAs were quantified by RT-PCR and active MMPs were quantified by ELISA.

Results: Our results show significantly (p<0.001) higher expression of CCR9 by mucinous adenocarcinoma, papillary serous carcinoma, and endometriod ovarian carcinoma cases, than compared to non-neoplastic ovarian tissue. Furthermore, CCR9 expression was significantly elevated in OvCa cell lines (OVCAR-3 and CAOV-3) in comparison to normal adult ovarian epithelial cell mRNA. OvCa cells showed higher migratory and invasive potential towards chemotactic gradients of CCL25, which was inhibited by anti-CCR9 antibodies. Expression of collagenases (MMP-1, -8, and -13), gelatinases (MMP-2 and -9), and stromelysins (MMP-3, -10, and -11) by OvCa cells were modulated by CCL25 in a CCR9-dependent fashion.

Conclusions: These results demonstrate both biological significance and clinical relevance of CCL25 and CCR9 interactions in OvCa cell metastasis.

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CCR9 expressed by ovarian cancer tissue. Ovarian cancer tissues from non- neoplastic (n = 8), mucinous adenocarcinoma (n = 5), serous papillary carcinoma (n = 10), and endometroid carcinoma (n = 11) were stained with isotype control or anti-CCR9 antibodies. Brown (DAB) color show CCR9 staining. An Aperio ScanScope CS system with a 40× objective captured digital images of each slide. Representative cases are indicated and immuno-intensities of CCR9 were quantified using image analysis Aperio ImageScope v.6.25 software. CCR9 expression by tissues were analyzed and presented by modified box plot. Lower, middle and upper lines, respectively, in the box represent the first quartile (Q1), Median (Q2) and third quartile (Q3). Upper (T) and lower (⊥) whiskers are represented by median ± 1.5 (Q3-Q1). Significant differences from non-neoplastic are indicated with a solid star whereas significant differences between mucinous adenocarcinoma and serous papillary as well as endometroid carcinomas are indicated with a white star in a black circle.
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Figure 1: CCR9 expressed by ovarian cancer tissue. Ovarian cancer tissues from non- neoplastic (n = 8), mucinous adenocarcinoma (n = 5), serous papillary carcinoma (n = 10), and endometroid carcinoma (n = 11) were stained with isotype control or anti-CCR9 antibodies. Brown (DAB) color show CCR9 staining. An Aperio ScanScope CS system with a 40× objective captured digital images of each slide. Representative cases are indicated and immuno-intensities of CCR9 were quantified using image analysis Aperio ImageScope v.6.25 software. CCR9 expression by tissues were analyzed and presented by modified box plot. Lower, middle and upper lines, respectively, in the box represent the first quartile (Q1), Median (Q2) and third quartile (Q3). Upper (T) and lower (⊥) whiskers are represented by median ± 1.5 (Q3-Q1). Significant differences from non-neoplastic are indicated with a solid star whereas significant differences between mucinous adenocarcinoma and serous papillary as well as endometroid carcinomas are indicated with a white star in a black circle.

Mentions: Ovarian TMAs consisting of non-neoplastic, mucinous adenocarcinoma, papillary serous carcinoma, and endometriod carcinoma tissues were evaluated for CCR9 expression. Positive staining was classified as 1 (missing or weak expression), 2 (medium expression), or 3 (high expression). In general, OvCa tissues significantly (p < 0.001) expressed CCR9 compared to non-neoplastic tissue, as did papillary serous and endometroid carcinomas compared to mucinous adenocarcinoma (Figure 1). The highest expression of CCR9 was observed in endometriod carcinoma followed by papillary serous carcinomas. While CCR9 expression by mucinous adenocarcinoma was lower than endometriod and papillary serous carcinomas, these OvCa cases significantly (p < 0.001) expressed CCR9 compared to non-neoplastic ovarian tissue.


CCL25-CCR9 interaction modulates ovarian cancer cell migration, metalloproteinase expression, and invasion.

Johnson EL, Singh R, Singh S, Johnson-Holiday CM, Grizzle WE, Partridge EE, Lillard JW - World J Surg Oncol (2010)

CCR9 expressed by ovarian cancer tissue. Ovarian cancer tissues from non- neoplastic (n = 8), mucinous adenocarcinoma (n = 5), serous papillary carcinoma (n = 10), and endometroid carcinoma (n = 11) were stained with isotype control or anti-CCR9 antibodies. Brown (DAB) color show CCR9 staining. An Aperio ScanScope CS system with a 40× objective captured digital images of each slide. Representative cases are indicated and immuno-intensities of CCR9 were quantified using image analysis Aperio ImageScope v.6.25 software. CCR9 expression by tissues were analyzed and presented by modified box plot. Lower, middle and upper lines, respectively, in the box represent the first quartile (Q1), Median (Q2) and third quartile (Q3). Upper (T) and lower (⊥) whiskers are represented by median ± 1.5 (Q3-Q1). Significant differences from non-neoplastic are indicated with a solid star whereas significant differences between mucinous adenocarcinoma and serous papillary as well as endometroid carcinomas are indicated with a white star in a black circle.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2927595&req=5

Figure 1: CCR9 expressed by ovarian cancer tissue. Ovarian cancer tissues from non- neoplastic (n = 8), mucinous adenocarcinoma (n = 5), serous papillary carcinoma (n = 10), and endometroid carcinoma (n = 11) were stained with isotype control or anti-CCR9 antibodies. Brown (DAB) color show CCR9 staining. An Aperio ScanScope CS system with a 40× objective captured digital images of each slide. Representative cases are indicated and immuno-intensities of CCR9 were quantified using image analysis Aperio ImageScope v.6.25 software. CCR9 expression by tissues were analyzed and presented by modified box plot. Lower, middle and upper lines, respectively, in the box represent the first quartile (Q1), Median (Q2) and third quartile (Q3). Upper (T) and lower (⊥) whiskers are represented by median ± 1.5 (Q3-Q1). Significant differences from non-neoplastic are indicated with a solid star whereas significant differences between mucinous adenocarcinoma and serous papillary as well as endometroid carcinomas are indicated with a white star in a black circle.
Mentions: Ovarian TMAs consisting of non-neoplastic, mucinous adenocarcinoma, papillary serous carcinoma, and endometriod carcinoma tissues were evaluated for CCR9 expression. Positive staining was classified as 1 (missing or weak expression), 2 (medium expression), or 3 (high expression). In general, OvCa tissues significantly (p < 0.001) expressed CCR9 compared to non-neoplastic tissue, as did papillary serous and endometroid carcinomas compared to mucinous adenocarcinoma (Figure 1). The highest expression of CCR9 was observed in endometriod carcinoma followed by papillary serous carcinomas. While CCR9 expression by mucinous adenocarcinoma was lower than endometriod and papillary serous carcinomas, these OvCa cases significantly (p < 0.001) expressed CCR9 compared to non-neoplastic ovarian tissue.

Bottom Line: OvCa tissue microarrays (TMA) was used to confirm CCR9 expression in clinical samples.Our results show significantly (p<0.001) higher expression of CCR9 by mucinous adenocarcinoma, papillary serous carcinoma, and endometriod ovarian carcinoma cases, than compared to non-neoplastic ovarian tissue.Expression of collagenases (MMP-1, -8, and -13), gelatinases (MMP-2 and -9), and stromelysins (MMP-3, -10, and -11) by OvCa cells were modulated by CCL25 in a CCR9-dependent fashion.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Biochemistry, & Immunology, Morehouse School of Medicine, 720 Westview Drive SW, Atlanta, GA 30310-1495, USA.

ABSTRACT

Background: Ovarian carcinoma (OvCa) is the most lethal gynecological malignancy among women and its poor prognosis is mainly due to metastasis. Chemokine receptor CCR9 is primarily expressed by a small subset of immune cells and its only natural ligand, CCL25, is largely expressed in the thymus, which involutes with age. Other than the thymus, CCL25 is expressed by the small bowel. Interactions between CCL25 and CCR9 have been implicated in leukocyte trafficking to the small bowel, a frequent metastatic site for OvCa cells. The current study shows OvCa tissue and cells significantly express CCR9, which interacts with CCL25 to support carcinoma cell migration and invasion.

Methods: RT-PCR and flow cytometry techniques were used to quantify the expression CCR9 by OvCa cells. OvCa tissue microarrays (TMA) was used to confirm CCR9 expression in clinical samples. The Aperio ScanScope scanning system was used to quantify immunohistochemical staining. Cell invasion and migration assays were performed using cell migration and matrigel invasion chambers. Matrix metalloproteinase (MMP) mRNAs were quantified by RT-PCR and active MMPs were quantified by ELISA.

Results: Our results show significantly (p<0.001) higher expression of CCR9 by mucinous adenocarcinoma, papillary serous carcinoma, and endometriod ovarian carcinoma cases, than compared to non-neoplastic ovarian tissue. Furthermore, CCR9 expression was significantly elevated in OvCa cell lines (OVCAR-3 and CAOV-3) in comparison to normal adult ovarian epithelial cell mRNA. OvCa cells showed higher migratory and invasive potential towards chemotactic gradients of CCL25, which was inhibited by anti-CCR9 antibodies. Expression of collagenases (MMP-1, -8, and -13), gelatinases (MMP-2 and -9), and stromelysins (MMP-3, -10, and -11) by OvCa cells were modulated by CCL25 in a CCR9-dependent fashion.

Conclusions: These results demonstrate both biological significance and clinical relevance of CCL25 and CCR9 interactions in OvCa cell metastasis.

Show MeSH
Related in: MedlinePlus