Influence of Paclitaxel and Doxorubicin Therapy of ßIII-Tubulin, Carbonic Anhydrase IX, and Survivin in Chemically Induced Breast Cancer in Female Rat

Breast cancer is the most common cancer in females. The aim of this study was to determine the effect of paclitaxel (PTX) and doxorubicin (DOX) therapy on the βIII-tubulin, carbonic anhydrase IX (CA IX), and survivin expression in chemically-induced rat mammary tumors. Animals with induced mammary carcinogenesis were randomly divided into treatment groups and an untreated group. The total proportion of tumors, the proportion of carcinoma in situ (CIS), and invasive carcinoma (IC) were evaluated. Protein expression in tumor tissue was determined using IHC. Statistical analysis of the data, evaluated by Fisher-exact test and unpaired t-test. Significantly increased levels of proteins in the tumor cells were confirmed using the IHC method for all studied proteins. The expression of βIII-tubulin, CA IX, and survivin increased significantly after treatment with both cytostatics (PTX and DOX). Depending on the type of tumor, a significant increase in all proteins was observed in IC samples after PTX treatment, and CA IX expression after DOX treatment. In CIS samples, a significant increase of βIII-tubulin and survivin expression was observed after a DOX treatment. The results suggest that βIII-tubulin, survivin, and CA IX may be significant drug resistance markers and the clinical regulation of their activity may be an effective means of reversing this resistance.

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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.

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Prognostic Significance of a Novel Hypoxia-Regulated Marker, Carbonic Anhydrase IX, in Invasive Breast Carcinoma

Journal of Clinical Oncology ◽

10.1200/jco.2001.19.16.3660 ◽

2001 ◽

Vol 19 (16) ◽

pp. 3660-3668 ◽

Cited By ~ 294

Author(s):

Stephen K. Chia ◽

Charles C. Wykoff ◽

Peter H. Watson ◽

Cheng Han ◽

Russell D. Leek ◽

...

Keyword(s):

Breast Cancer ◽

Overall Survival ◽

Breast Carcinoma ◽

Carbonic Anhydrase ◽

Invasive Breast Cancer ◽

Estrogen Receptor Status ◽

Prognostic Significance ◽

Invasive Breast Carcinoma ◽

Carbonic Anhydrase Ix ◽

Ca Ix

PURPOSE: To assess the frequency of expression and the prognostic significance of a hypoxia-regulated marker, carbonic anhydrase IX (CA IX), in a cohort of patients with invasive breast cancer. PATIENTS AND METHODS: CA IX expression was evaluated by immunohistochemistry with a murine monoclonal antibody, M75, in a series of 103 women treated surgically for invasive breast cancer. The majority of patients were treated with adjuvant hormonal or chemotherapy. The frequency of CA IX expression, its association with recognized prognostic factors, and the relationship with outcome was evaluated by univariate and multivariate statistical analyses. RESULTS: CA IX expression was present in 49 (48%) of 103 cases. The level of CA IX expression was found to be significantly associated with tumor necrosis (P < .001), higher grade (P = .02), and negative estrogen receptor status (P < .001). Furthermore, CA IX expression was associated with a higher relapse rate (P = .004) and a worse overall survival (P = .001). By multivariate analysis, CA IX was also shown to be an independent predictive factor for overall survival (hazard ratio, 2.61; 95% confidence interval, 1.01 to 6.75, P = .05). CONCLUSION: CA IX expression was associated with worse relapse-free survival and overall survival in an unselected cohort of patients with invasive breast carcinoma. The potential role of CA IX as a marker of hypoxia within breast carcinomas was also indicated by a significant association with necrosis. Further work assessing its prognostic significance in breast cancer is warranted, particularly interactions with radiotherapy and chemotherapy resistance.

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770 poster PROGNOSTIC SIGNIFICANCE OF CARBONIC ANHYDRASE IX (CA IX) IN BREAST CANCER PATIENTS TREATED WITH RADIOTHERAPY

Radiotherapy and Oncology ◽

10.1016/s0167-8140(11)70892-1 ◽

2011 ◽

Vol 99 ◽

pp. S303-S304

Author(s):

L. Beketic-Oreskovic ◽

P. Ozretic ◽

Z. Rabbani ◽

I. Jackson ◽

B. Sarcevic ◽

...

Keyword(s):

Breast Cancer ◽

Carbonic Anhydrase ◽

Cancer Patients ◽

Prognostic Significance ◽

Carbonic Anhydrase Ix ◽

Breast Cancer Patients ◽

Ca Ix

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Prognostic Significance of Carbonic Anhydrase IX (CA-IX), Endoglin (CD105) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in Breast Cancer Patients

Pathology & Oncology Research ◽

10.1007/s12253-010-9355-6 ◽

2011 ◽

Vol 17 (3) ◽

pp. 593-603 ◽

Cited By ~ 20

Author(s):

Lidija Beketic-Oreskovic ◽

Petar Ozretic ◽

Zahid N. Rabbani ◽

Isabel L. Jackson ◽

Bozena Sarcevic ◽

...

Keyword(s):

Breast Cancer ◽

Carbonic Anhydrase ◽

Cancer Patients ◽

Prognostic Significance ◽

Carbonic Anhydrase Ix ◽

Breast Cancer Patients ◽

Ca Ix

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

10.1101/505644 ◽

2018 ◽

Cited By ~ 1

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 cooperates 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 (TNBC), appears to coordinately 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.

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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.

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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.

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Granulocyte Colony Stimulating Factor Expression in Breast Cancer and Its Association with Carbonic Anhydrase IX and Immune Checkpoints

Cancers ◽

10.3390/cancers13051022 ◽

2021 ◽

Vol 13 (5) ◽

pp. 1022

Author(s):

Shawn C. Chafe ◽

Nazia Riaz ◽

Samantha Burugu ◽

Dongxia Gao ◽

Samuel C. Y. Leung ◽

...

Keyword(s):

Breast Cancer ◽

Carbonic Anhydrase ◽

Immune Checkpoint ◽

Positive Association ◽

Carcinoma Cells ◽

Carbonic Anhydrase Ix ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Significant Positive Association ◽

Stimulating Factor

Purpose: Granulocyte colony-stimulating factor (G-CSF) and hypoxia modulate the tumour immune microenvironment. In model systems, hypoxia-induced carbonic anhydrase IX (CAIX) has been associated with G-CSF and immune responses, including M2 polarization of macrophages. We investigated whether these associations exist in human breast cancer specimens, their relation to breast cancer subtypes, and clinical outcome. Methods: Using validated protocols and prespecified scoring methodology, G-CSF expression on carcinoma cells and CD163 expression on tumour-associated macrophages were assayed by immunohistochemistry and applied to a tissue microarray series of 2960 primary excision specimens linked to clinicopathologic, biomarker, and outcome data. Results: G-CSFhigh expression showed a significant positive association with ER negativity, HER2 positivity, presence of CD163+ M2 macrophages, and CAIX expression. In univariate analysis, G-CSFhigh phenotype was associated with improved survival in non-luminal cases, although the CAIX+ subset had a significantly adverse prognosis. A significant positive association was observed between immune checkpoint biomarkers on tumour-infiltrating lymphocytes and both G-CSF- and CAIX-expressing carcinoma cells. Immune checkpoint biomarkers correlated significantly with favourable prognosis in G-CSFhigh/non-luminal cases independent of standard clinicopathological features. Conclusions: The prognostic associations linking G-CSF to immune biomarkers and CAIX strongly support their immunomodulatory roles in the tumour microenvironment.

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