Carbonic anhydrase 9 (CA9) expression in tumor cells enhances sensitivity to tirapazamine

Shin, Hyung Shik; Kim, Joo-Young; Yoo, Chong-Woo; Roberts, Stephen A.; Lee, Sun; Choi, Soo‐Jin; Lee, Heeyoung; Lee, Doo-Hyun; Kim, Tae Hyun; Cho, Kwan Ho

doi:10.1007/s00432-007-0298-6

Cited by 5 publications

(8 citation statements)

References 25 publications

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“…This pattern was very similar to the morphological changes of HeLa cells in which CA9-expression was greatly stimulated by hypoxic conditions (Fig. 2B) (Shin et al, 2008). The expression and tyrosine phosphorylation of paxillin, a major regulator in organization and function of focal adhesions, were assessed by western blot analysis (Fig.…”

Section: Effect Of Ca9 On Formation Of Actin Stress Fiber and Focal Asupporting

confidence: 81%

“…The human cervical cancer cell line C33A (ATCC number HTB-31) was stably transfected with either full-length human CA9 cDNA cloned into the pcDNA3 vector or empty vector control (Shin et al, 2008). Transfected polyclonal cell populations were selected for growth in the presence of 250 g/ml G418 (GIBCO-BRL, Gaithersburg, MD) for 3 weeks.…”

Section: Cell Culture and Stable Transfection Of C33a Cellsmentioning

confidence: 99%

“…Transfected polyclonal cell populations were selected for growth in the presence of 250 g/ml G418 (GIBCO-BRL, Gaithersburg, MD) for 3 weeks. Selected monoclonal CA9-transfected cells (C33A/CA9) and vector-only-transfected cells (C33A/Mock) were tested for expression of CA9 as previously described by real-time PCR and western blot analysis (Shin et al, 2008). M75, the monoclonal antibody for CA9, was a kind gift from Silvia Pastorekova (Slovak Academy of Sciences, Slovakia).…”

Section: Cell Culture and Stable Transfection Of C33a Cellsmentioning

confidence: 99%

“…To search for the key regulators of metastasis related to CA9 expression under hypoxic conditions, we established a cell line that constitutively expresses CA9 using the C33A human cervical cancer cell line (Shin et al, 2008). We then further investigated differentially expressed gene profiles in CA9-transfected C33A cervix cancer cells (C33A/CA9) compared with vector-onlytransfected cells (C33A/Mock) (Shin et al, 2008), using the Affymetrix Human Genome U133 Plus gene chip assay.…”

Section: Introductionmentioning

confidence: 99%

“…We then further investigated differentially expressed gene profiles in CA9-transfected C33A cervix cancer cells (C33A/CA9) compared with vector-onlytransfected cells (C33A/Mock) (Shin et al, 2008), using the Affymetrix Human Genome U133 Plus gene chip assay. Because our previous studies consistently revealed the high metastatic potential in tumors where CA9 is overexpressed (Kim et al, 2006;Lee et al, 2007), we focused on the gene ontology categories related to metastatic phenotypes.…”

Section: Introductionmentioning

confidence: 99%

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Carbonic anhydrase IX (CA9) modulates tumor-associated cell migration and invasion

Shin

Rho

Jung

et al. 2011

Journal of Cell Science

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109

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SummaryExpression of carbonic anhydrase IX (CA9) was shown to be strongly involved in high incidences of metastasis and poor prognosis in various human tumors. In this study, we investigated the possible role for CA9 in tumor metastases in vitro, using a gene transfection tool in the human cervical carcinoma cell line C33A. Gene expression profiling of CA9-transfected cells (C33A/CA9) and vectortransfected cells (C33A/Mock) was investigated by DNA microarray. The biological functions of differentially expressed genes between the C33A/CA9 and C33A/Mock cells included cell growth, regulation of cell-cell and cell-extracellular matrix adhesion and cytoskeletal organization. Immunofluorescent stain and Matrigel culture showed cytoskeletal remodeling, disassembled focal adhesion, weakened cell-cell adhesion and increased motility in C33A/CA9 cells. These invasive and metastatic phenotypes were associated with Rho-GTPase-related epithelial-mesenchymal transition. Inhibition of the Rho/Rho kinase pathway by a ROCK inhibitor (Y27632) and si-Rho (short interference RNA against RhoA) showed that Rho-GTPase signaling was involved in cellular morphologic and migratory changes. The effect of CA9 on Rho-GTPase signaling was also confirmed by silencing CA9 expression. Our results suggest that CA9 overexpression induces weakening of cell adhesions and augmented cell motility by aberrant Rho-GTPase signal transduction. Our study shows an underlying mechanism of CA9-related enhanced metastatic potential of tumor cells.

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Section: Effect Of Ca9 On Formation Of Actin Stress Fiber and Focal Asupporting

confidence: 81%

Section: Cell Culture and Stable Transfection Of C33a Cellsmentioning

confidence: 99%

Section: Cell Culture and Stable Transfection Of C33a Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Carbonic anhydrase IX (CA9) modulates tumor-associated cell migration and invasion

Shin

Rho

Jung

et al. 2011

Journal of Cell Science

Self Cite

109

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Hypoxia-targeting by tirapazamine (TPZ) induces preferential growth inhibition of nasopharyngeal carcinoma cells with Chk1/2 activation

et al. 2009

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Hypoxia is commonly developed in solid tumors, which contributes to metastasis as well as radio- and chemo-resistance. Nasopharyngeal carcinoma (NPC) is a highly invasive and metastatic head and neck cancer prevalent in Southeast Asia with a high incidence rate of 15-30/100,000 persons/year (comparable to that of pancreatic cancer in the US). Previous clinical studies in NPC showed that hypoxia is detected in almost 100% of primary tumors and overexpression of hypoxia markers correlated with poor clinical outcome. Tirapazamine (TPZ) is a synthetic hypoxia-activated prodrug, which preferentially forms cytotoxic and DNA-damaging free radicals under hypoxia, thus selectively eradicate hypoxic cells. Here, we hypothesized that specific hypoxia-targeting by this clinical trial agent may be therapeutic for NPC. Our findings demonstrated that under hypoxia, TPZ was able to induce preferential growth inhibition of NPC cells, which was associated with marked cell cycle arrest at S-phase and PARP cleavage (a hallmark of apoptosis). Examination of S-phase checkpoint regulators revealed that Chk1 and Chk2 were selectively activated by TPZ in NPC cells under hypoxia. Hypoxia-selectivity of TPZ was also demonstrated by preferential downregulation of several important hypoxia-induced markers (HIF-1α, CA IX and VEGF) under hypoxia. Furthermore, we demonstrated that TPZ was equally effective and hypoxia-selective even in the presence of the EBV oncoprotein, LMP1 or the EBV genome. In summary, encouraging results from this proof-of-concept study implicate the therapeutic potential of hypoxia-targeting approaches for the treatment of NPC.

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Current World Literature

2009

Current Opinion in Otolaryngology &Amp; Head &Amp; Neck Surgery

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Carbonic anhydrase 9 (CA9) expression in tumor cells enhances sensitivity to tirapazamine

Abstract: Our results suggest the possibility that CA9 over-expression in tumors might be exploited to increase the treatment effects of TPZ.

Cited by 5 publications

References 25 publications

Carbonic anhydrase IX (CA9) modulates tumor-associated cell migration and invasion

Carbonic anhydrase IX (CA9) modulates tumor-associated cell migration and invasion

Hypoxia-targeting by tirapazamine (TPZ) induces preferential growth inhibition of nasopharyngeal carcinoma cells with Chk1/2 activation

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