Immunohistochemical Study of Colorectal Tumors for Expression of a Novel Transmembrane Carbonic Anhydrase, MN/CA IX, with Potential Value as a Marker of Cell Proliferation

Object Carbonic anhydrase (CA) II and IX are enzymes involved in pH homeostasis and have been shown to be upregulated in several types of cancer. In this study, the authors evaluate the expression of CA II and IX in meningiomas and assess their relationship to patient age, tumor type and grade, tumor sex hormone receptor status, tumor cell proliferation, and tumor recurrence. Methods This study was conducted in consecutive patients who underwent meningioma surgeries at Tampere University Hospital between 1989 and 1999. The expression of CA II and IX was studied immunohistochemically using a tissue microarray technique and specific antibodies. Results Immunohistological staining with CA II and IX was assessed in 443 primary and 67 recurrent tumor specimens. Of these samples, 455 were benign (WHO Grade I), 49 atypical (Grade II), and 6 malignant (Grade III). Endothelial cells in 14.8% of the tumors stained positively for CA II. Tumor cells were positive for CA IX in 11.6% of the cases. Endothelial CA II expression correlated with increasing histological grade (p = 0.002), and tumor proliferation rates were higher in CA II+ versus CA II− cases (p = 0.002). Androgen receptor–negative tumors were found to be CA II+ significantly more often than androgen receptor–positive tumors (p = 0.001). No associations were found with the CA IX enzyme. Conclusions Carbonic anhydrase II positivity in the endothelium was associated with cell proliferation and malignancy grade. These results suggest that CA II expression is associated with malignant progression of meningiomas and could thus be a target molecule for anticancer therapy.

Download Full-text

Immunohistochemistry of Carbonic Anhydrase Isozyme IX (MN/CA IX) in Human Gut Reveals Polarized Expression in the Epithelial Cells with the Highest Proliferative Capacity

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215549804600409 ◽

1998 ◽

Vol 46 (4) ◽

pp. 497-504 ◽

Cited By ~ 105

Author(s):

Juha Saarnio ◽

Seppo Parkkila ◽

Anna-Kaisa Parkkila ◽

Abdul Waheed ◽

Matthew C. Casey ◽

...

Keyword(s):

Cell Proliferation ◽

Epithelial Cells ◽

Carbonic Anhydrase ◽

Plasma Membranes ◽

Human Gut ◽

Ileal Mucosa ◽

Western Blots ◽

Nih3t3 Cells ◽

Ca Ix ◽

Specific Regulation

MN/CA IX is a recently discovered member of the carbonic anhydrase (CA) gene family that has been identified in the plasma membranes of certain tumor and epithelial cells and found to promote cell proliferation when transfected into NIH3T3 cells. This study presents localization of MN/CA IX in human gut and compares its distribution to those of CA I, II, and IV, which are known to be expressed in the intestinal epithelium. The specificity of the monoclonal antibody for MN/CA IX was confirmed by Western blots and immunostaining of COS-7 cells transfected with MN/CA IX cDNA. Immunohistochemical stainings of human gut revealed prominent polarized staining for MN/CA IX in the basolateral surfaces of the enterocytes of duodenum and jejunum, the reaction being most intense in the crypts. A moderate reaction was also seen in the crypts of ileal mucosa, whereas the staining became generally weaker in the large intestine. The results indicate isozyme-specific regulation of MN/CA IX expression along the cranial–caudal axis of the human gut and place the protein at the sites of rapid cell proliferation. The unique localization of MN/CA IX on the basolateral surfaces of proliferating crypt enterocytes suggests that it might serve as a ligand or a receptor for another protein that regulates intercellular communication or cell proliferation. Furthermore, MN/CA IX has a completely conserved active site domain of CAs suggesting that it could also participate in carbon dioxide/bicarbonate homeostasis.

Download Full-text

Distinct Expression Patterns of Carbonic Anhydrase IX in Clear Cell, Microcystic, and Angiomatous Meningiomas

Journal of Neuropathology & Experimental Neurology ◽

10.1093/jnen/nlz091 ◽

2019 ◽

Vol 78 (12) ◽

pp. 1081-1088

Author(s):

Rati Chkheidze ◽

Patrick J Cimino ◽

Kimmo J Hatanpaa ◽

Charles L White ◽

Manuel Ferreira ◽

...

Keyword(s):

Carbonic Anhydrase ◽

Clear Cell ◽

Expression Patterns ◽

Carbonic Anhydrase Ix ◽

World Health ◽

Loss Of Function ◽

Ca Ix ◽

Hypoxia Marker ◽

Other World ◽

Health Organization

Abstract Clear cell, microcytic, and angiomatous meningiomas are 3 vasculature-rich variants with overlapping morphological features but different prognostic and treatment implications. Distinction between them is not always straightforward. We compared the expression patterns of the hypoxia marker carbonic anhydrase IX (CA-IX) in meningiomas with predominant clear cell (n = 15), microcystic (n = 9), or angiomatous (n = 11) morphologies, as well as 117 cases of other World Health Organization recognized histological meningioma variants. Immunostaining for SMARCE1 protein, whose loss-of-function has been associated with clear cell meningiomas, was performed on all clear cell meningiomas, and selected variants of meningiomas as controls. All clear cell meningiomas showed absence of CA-IX expression and loss of nuclear SMARCE1 expression. All microcystic and angiomatous meningiomas showed diffuse CA-IX immunoreactivity and retained nuclear SMARCE1 expression. In other meningioma variants, CA-IX was expressed in a hypoxia-restricted pattern and was highly associated with atypical features such as necrosis, small cell change, and focal clear cell change. In conclusion, CA-IX may serve as a useful diagnostic marker in differentiating clear cell, microcystic, and angiomatous meningiomas.

Download Full-text

Inhibition of Carbonic Anhydrase IX Promotes Apoptosis through Intracellular pH Level Alterations in Cervical Cancer Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22116098 ◽

2021 ◽

Vol 22 (11) ◽

pp. 6098

Author(s):

Ebru Temiz ◽

Ismail Koyuncu ◽

Mustafa Durgun ◽

Murat Caglayan ◽

Ataman Gonel ◽

...

Keyword(s):

Carbonic Anhydrase ◽

Intracellular Ph ◽

Hela Cells ◽

Protein Level ◽

Caspase 3 ◽

Solid Tumours ◽

Carbonic Anhydrase Ix ◽

Parp Activation ◽

Ca Ix ◽

Cervical Cancer Cells

Carbonic anhydrase IX (CAIX) is a hypoxia-related protein that plays a role in proliferation in solid tumours. However, how CAIX increases proliferation and metastasis in solid tumours is unclear. The objective of this study was to investigate how a synthetic CAIX inhibitor triggers apoptosis in the HeLa cell line. The intracellular effects of CAIX inhibition were determined with AO/EB, AnnexinV-PI, and γ-H2AX staining; measurements of intracellular pH (pHi), reactive oxygen species (ROS), and mitochondrial membrane potential (MMP); and analyses of cell cycle, apoptotic, and autophagic modulator gene expression (Bax, Bcl-2, caspase-3, caspase-8, caspase-9, caspase-12, Beclin, and LC3), caspase protein level (pro-caspase 3 and cleaved caspase-3, -8, -9), cleaved PARP activation, and CAIX protein level. Sulphonamide CAIX inhibitor E showed the lowest IC50 and the highest selectivity index in CAIX-positive HeLa cells. CAIX inhibition changed the morphology of HeLa cells and increased the ratio of apoptotic cells, dramatically disturbing the homeostasis of intracellular pHi, MMP and ROS levels. All these phenomena consequent to CA IX inhibition triggered apoptosis and autophagy in HeLa cells. Taken together, these results further endorse the previous findings that CAIX inhibitors represent an important therapeutic strategy, which is worth pursuing in different cancer types, considering that presently only one sulphonamide inhibitor, SLC-0111, has arrived in Phase Ib/II clinical trials as an antitumour/antimetastatic drug.

Download Full-text

1,2-Benzisothiazole Derivatives Bearing 4-, 5-, or 6-Alkyl/arylcarboxamide Moieties Inhibit Carbonic Anhydrase Isoform IX (CAIX) and Cell Proliferation under Hypoxic Conditions

Journal of Medicinal Chemistry ◽

10.1021/acs.jmedchem.6b00616 ◽

2016 ◽

Vol 59 (13) ◽

pp. 6547-6552 ◽

Cited By ~ 13

Author(s):

Vito Coviello ◽

Beatrice Marchi ◽

Stefania Sartini ◽

Luca Quattrini ◽

Anna Maria Marini ◽

...

Keyword(s):

Cell Proliferation ◽

Carbonic Anhydrase ◽

Hypoxic Conditions

Download Full-text

Immunohistochemical study of carbonic anhydrase in mixed tumours from major salivary glands and skin

Virchows Archiv A Pathological Anatomy and Histopathology ◽

10.1007/bf00705299 ◽

1986 ◽

Vol 408 (5) ◽

pp. 449-459 ◽

Cited By ~ 15

Author(s):

Yohko Noda ◽

Yoshiaki Takai ◽

Yoshimasa Iwai ◽

Michael A. Meenaghan ◽

Masahiko Mori

Keyword(s):

Carbonic Anhydrase ◽

Salivary Glands ◽

Immunohistochemical Study ◽

Major Salivary Glands ◽

Mixed Tumours

Download Full-text

Carbonic anhydrase IX enhances tumor cell proliferation and tumor progression in osteosarcoma

OncoTargets and Therapy ◽

10.2147/ott.s177605 ◽

2018 ◽

Vol Volume 11 ◽

pp. 6879-6886 ◽

Cited By ~ 6

Author(s):

Kazuma Okuno ◽

Takao Matsubara ◽

Tomoki Nakamura ◽

Takahiro Iino ◽

Takuya Kakimoto ◽

...

Keyword(s):

Cell Proliferation ◽

Carbonic Anhydrase ◽

Tumor Cell ◽

Tumor Progression ◽

Carbonic Anhydrase Ix ◽

Tumor Cell Proliferation

Download Full-text

Immunohistochemical Study on Apoptosis and Cell Proliferation in the Lining Epithelium of Thyroglossal Duct Cysts.

Oral Medicine & Pathology ◽

10.3353/omp.4.47 ◽

1999 ◽

Vol 4 (2) ◽

pp. 47-54 ◽

Cited By ~ 1

Author(s):

Mitsuhide Yoshida ◽

Hiroyuki Kumamoto ◽

Kiyoshi Ooya ◽

Hideaki Mayanagi

Keyword(s):

Cell Proliferation ◽

Immunohistochemical Study ◽

Thyroglossal Duct ◽

Thyroglossal Duct Cysts

Download Full-text

Comparative Evaluation of the Clinical Efficacy of PRP-Therapy, Minoxidil, and Their Combination with Immunohistochemical Study of the Dynamics of Cell Proliferation in the Treatment of Men with Androgenetic Alopecia

International Journal of Molecular Sciences ◽

10.3390/ijms21186516 ◽

2020 ◽

Vol 21 (18) ◽

pp. 6516

Author(s):

Elena E. Pakhomova ◽

Irina O. Smirnova

Keyword(s):

Cell Proliferation ◽

Clinical Efficacy ◽

Comparative Evaluation ◽

Proliferative Activity ◽

Immunohistochemical Study ◽

Platelet Rich Plasma ◽

Androgenetic Alopecia ◽

Hair Morphology ◽

Promising Treatment ◽

Cd34 Expression

Platelet-rich plasma (PRP) therapy has been considered as a promising treatment for androgenetic alopecia (AGA). The aim of the study was comparative evaluation of the clinical efficacy of PRP-therapy, minoxidil, and their combination in the treatment of men with AGA and to evaluate the effects of PRP on the proliferation of hair follicle (HF) cells in skin biopsy. Materials and Methods: The study involved 69 men who were divided into 3 groups who received PRP therapy, minoxidil, and their combination. The clinical efficacy of the therapy was evaluated by the dynamics of morphometric of hairs. To assess cell proliferation antibodies to β-catenin, CD34, Ki67, and to Dkk-1 were used. Results. PRP treatment was more effective than minoxidil therapy (p = 0.005). Complex therapy turned out to be more effective than minoxidil monotherapy (p < 0.0001) and PRP monotherapy (p = 0.007). After applying PRP the absolute and relative values of the β-catenin and CD34 expression area increased; an increase in Ki67+ index was also significant. Conclusions: PRP can be considered as a treatment option for AGA. Combined PRP and minoxidil use seems promising for the treatment of AGA. PRP increase in the proliferative activity of HF cells and improves hair morphology in patients with AGA.

Download Full-text

A non-catalytic function of carbonic anhydrase IX contributes to the glycolytic phenotype and pH regulation in human breast cancer cells

Biochemical Journal ◽

10.1042/bcj20190177 ◽

2019 ◽

Vol 476 (10) ◽

pp. 1497-1513 ◽

Cited By ~ 8

Author(s):

Mam Y. Mboge ◽

Zhijuan Chen ◽

Daniel Khokhar ◽

Alyssa Wolff ◽

Lingbao Ai ◽

...

Keyword(s):

Breast Cancer ◽

Carbonic Anhydrase ◽

Cancer Cells ◽

Cancer Progression ◽

Expression Patterns ◽

Extracellular Ph ◽

Carbonic Anhydrase Ix ◽

Monocarboxylate Transporter ◽

Proton Efflux ◽

Ca Ix

AbstractThe most aggressive and invasive tumor cells often reside in hypoxic microenvironments and rely heavily on rapid anaerobic glycolysis for energy production. This switch from oxidative phosphorylation to glycolysis, along with up-regulation of the glucose transport system, significantly increases the release of lactic acid from cells into the tumor microenvironment. Excess lactate and proton excretion exacerbate extracellular acidification to which cancer cells, but not normal cells, adapt. We have hypothesized that carbonic anhydrases (CAs) play a role in stabilizing both intracellular and extracellular pH to favor cancer progression and metastasis. Here, we show that proton efflux (acidification) using the glycolytic rate assay is dependent on both extracellular pH (pHe) and CA IX expression. Yet, isoform-selective sulfonamide-based inhibitors of CA IX did not alter proton flux, which suggests that the catalytic activity of CA IX is not necessary for this regulation. Other investigators have suggested the CA IX co-operates with the MCT transport family to excrete protons. To test this possibility, we examined the expression patterns of selected ion transporters and show that members of this family are differentially expressed within the molecular subtypes of breast cancer. The most aggressive form of breast cancer, triple-negative breast cancer, appears to co-ordinately express the monocarboxylate transporter 4 (MCT4) and carbonic anhydrase IX (CA IX). This supports a possible mechanism that utilizes the intramolecular H+ shuttle system in CA IX to facilitate proton efflux through MCT4.

Download Full-text