Cytotoxic effects of adenovirus- and lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase on Bcap37 breast cancer cells

Zhang, Nianqu; Dong, Xinhua; Sun, Yiqun; Cai, Xiaopeng; Zheng, Caiwei; He, Anning; Xu, Ke; Zheng, Xinyu

doi:10.3892/or.2012.2194

Cited by 8 publications

(8 citation statements)

References 26 publications

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“…The truncated variant A significant increase in the sensitivity of transfected cell lines to gemcitabine was observed, as shown by a decrease in IC 50 compared with control group. The IC 50 and folds of higher than previously reported for human uterine sarcoma cell line Messa 10K, where a full-length DmdNK was transfected to sensitize human cancer cell lines to various nucleosides [9,[41][42][43][50][51][52]. A possible explanation for the increased sensitivity to gemcitabine is the lower K m value of DmdNKΔC20 for this analog and unique interaction of fluoride atoms of gemcitabine [40].…”

Section: Discussionmentioning

confidence: 76%

“…DmdNK and its improved variants, produced by site-directed mutagenesis, have been tested for their optimum application in suicide gene therapy [38][39][40]. Both viral (adeno-and lentivirus-based vectors) and non-viral delivery systems have been used for transfecting different cancer cells with DmdNK and its mutants, to study the combined cytotoxic effect of gene/prodrug and to reverse drug resistance in various cancer cell lines [9,[41][42][43]. To the best of our knowledge, DmdNKΔC20 has not been reported neither for sensitizing human cancer cell lines to gemcitabine nor for the reversal of gemcitabine resistance in drug-resistant cancer cells so far.…”

Section: Introductionmentioning

confidence: 99%

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Recombinant deoxyribonucleoside kinase from Drosophila melanogaster can improve gemcitabine based combined gene/chemotherapy for targeting cancer cells

Fatima

Ahmed

Batool

et al. 2019

Bosn J of Basic Med Sci

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A recombinant deoxyribonucleoside kinase from Drosophila melanogaster with a deletion of the last 20 amino acid residues (named DmdNKΔC20) was hypothesized as a potential therapeutic tool for gene therapy due to its broad substrate specificity and better catalytic efficiency towards nucleosides and nucleoside analogs. This study was designed to evaluate the effect of DmdNKΔC20 for sensitizing human cancer cell lines towards gemcitabine and to further investigate its role in reversal of acquired drug resistance in gemcitabine-resistant cancer cell line. The DmdNKΔC20 gene was delivered to three different cancer cell lines, including breast, colon and liver cancer cells, using lipid-mediated transfection reagent. After transfection, gene expression of DmdNKΔC20 was confirmed by reverse transcription quantitative PCR (qRT-PCR) and the combined effect of DmdNKΔC20 and gemcitabine based cytotoxicity was observed by cell viability assay. We further evolved a gemcitabine-resistant breast cancer cell line (named MCF7-R) through directed evolution in the laboratory, which showed 375-fold more resistance compared to parental MCF7 cells. Upon transfection with DmdNKΔC20 gene, MCF7-R cells showed 83-fold higher sensitivity to gemcitabine compared to the control group of MCF7-R cells. Moreover, we observed 79% higher expression of p21 protein in transfected MCF7-R cells, which may indicate induction of apoptosis. Our findings highlight the importance and therapeutic potential of DmdNKΔC20 in combined gene/chemotherapy approach to target a wide range of cancers, particularly gemcitabine-resistant cancers.

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Section: Discussionmentioning

confidence: 76%

Section: Introductionmentioning

confidence: 99%

Recombinant deoxyribonucleoside kinase from Drosophila melanogaster can improve gemcitabine based combined gene/chemotherapy for targeting cancer cells

Fatima

Ahmed

Batool

et al. 2019

Bosn J of Basic Med Sci

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“…It is evident that expression of a single transgene is unlikely to be sufficient to eradicate cancer. Numerous studies have demonstrated that HSV-tk combined with cytokine therapy for the treatment of breast carcinoma had a high efficacy (18)(19)(20)(21). It has been shown that specific expression of CD or HSV-tk genes in breast as well as other cancer cells can be achieved using a vector delivery system containing VEGF promoter sequences (22).…”

Section: Discussionmentioning

confidence: 99%

Thymidine kinase/ganciclovir and cytosine deaminase/5-fluorocytosine suicide gene therapy-induced cell apoptosis in breast cancer cells

Kong

Qin

et al. 2013

Oncology Reports

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The present study was conducted to explore the efficacy of suicide gene therapy with thymidine kinase (TK) in combination with cytosine deaminase (CD) for breast cancer. The expression of CD/TK was detected in the infected cells by RT-PCR. The killing effect on MCF-7 cells following treatment was analyzed by MTT assay. The morphological characteristics of the cells were observed by electron microscopy, and the distribution of the cell cycle was analyzed by flow cytometry. Caspase‑3 and -8 activities were detected by absorption spectrometry. Cytotoxic assays showed that cells transfected with CD/TK became more sensitive to the prodrugs. Morphological features characteristic of apoptosis were noted in the MCF‑7 cells via electron microscopy. The experimental data showed that the proportion of MCF-7 cells during the different phases of the cell cycle varied significantly following treatment with the prodrugs. The activity of caspase‑3 gradually increased following treatment with increasing concentrations of the prodrugs. We conclude that the TK/ganciclovir and CD/5-fluorocytosine suicide gene system used here induces apoptosis in breast cancer cells, and provides a promising treatment modality for breast cancer.

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“…Notably, the lentiviral vector has been identified to facilitate the incorporation of target genes into the host genome for efficient and stable expression of target genes in dividing and non-dividing cells (11,12). In the present study, a lentivirus vector was used to express the Dm-dNK suicide gene in keloid fibroblasts (KF).…”

Section: Introductionmentioning

confidence: 99%

Efficacy of lentivirus‑mediated Drosophila�melanogaster deoxyribonucleoside kinase combined with (E)‑5‑(2‑bromovinyl)‑2'‑deoxyuridine or 1‑β‑D‑arabinofuranosylthymine therapy in human keloid fibroblasts

Sun¹,

Jiang

et al. 2018

Mol Med Report

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Keloid scarring is a type of fibroproliferative disease with a high recurrence rate. However, no effective treatment is currently available. Combined therapy with recombinant lentivirus‑mediated Drosophila melanogaster deoxyribonucleoside kinase (Dm‑dNK) and prodrug has been widely studied and used for cancer treatment. Due to the similarities between keloid scars and tumors, the aim of the present study was to investigate the efficacy of a Dm‑dNK/nucleoside analog system for the treatment of keloid scars. Recombinant lentivirus expression of the Dm‑dNK suicide gene was assessed. Western blotting was used to examine the protein expression of lentivirus mediated Dm‑dNK in keloid fibroblasts. Enzyme activity assays were conducted using [3H]‑labeled substrates. Furthermore, cytotoxicity and bystander effects were evaluated using MTT assays. The expression of green fluorescent protein was observed using fluorescence microscope and results indicated that there was no notable difference in lentivirus infectivity between the multiplicity of infection (MOI) of 1 and 10 in cells. Notably, western blotting revealed that Dm‑dNK was stably expressed in keloid fibroblasts and the enzymatic activity assays revealed that the enzyme was activated following introduction into the keloid fibroblasts via the lentivirus. The cytotoxicity and bystander effects of Dm‑dNK combined with cytotoxic nucleoside analogs were both observed in Dm‑dNK+ keloid fibroblasts. These results demonstrated that the lentivirus‑mediated Dm‑dNK therapy may be effective in treating keloid fibroblasts, which provides some evidence for the use of Dm‑dNK/prodrug therapy for keloid treatment in vivo in the future.

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Cytotoxic effects of adenovirus- and lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase on Bcap37 breast cancer cells

Cited by 8 publications

References 26 publications

Recombinant deoxyribonucleoside kinase from Drosophila melanogaster can improve gemcitabine based combined gene/chemotherapy for targeting cancer cells

Recombinant deoxyribonucleoside kinase from Drosophila melanogaster can improve gemcitabine based combined gene/chemotherapy for targeting cancer cells

Thymidine kinase/ganciclovir and cytosine deaminase/5-fluorocytosine suicide gene therapy-induced cell apoptosis in breast cancer cells

Efficacy of lentivirus‑mediated Drosophila�melanogaster deoxyribonucleoside kinase combined with (E)‑5‑(2‑bromovinyl)‑2'‑deoxyuridine or 1‑β‑D‑arabinofuranosylthymine therapy in human keloid fibroblasts

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