Flow-cytometric analysis of DNA distribution after VP16-213 treatment of Lewis lung carcinoma

Erba, Eugenio; Ubezio, Paolo; Colombo, Tina; Broggini, Massimo; Torti, L.; Vaghi, Massimo; D’Incalci, Maurizio; Morasca, L.

doi:10.1007/bf00255765

Cited by 17 publications

(3 citation statements)

References 11 publications

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“…bation of the cell cycle and of the cell growth inhibition Figure 5 shows the cell growth inhibition effect on caused by this DNA staining. The perturbation of the L1210 cells in vitro exposed for 2 h at different concen-cell cycle consisted in an accumulation of cells in premitrations of HO 33342 evaluated after a recovery time of totic phase, an effect already previously observed for 24,48,72, and 144 h. After 24 h, inhibition of cell growth other DNA-damaging agents (4). In addition HO 33342 was already evident, even at a concentration of 0.5 pg/ induced the appearance of tetraploid cell population ml, a concentration tenfold smaller than those necessary which was not detectable before treatment.…”

Section: Discussionmentioning

confidence: 71%

DNA damage, cytotoxic effect and cell‐cycle perturbation of Hoechst 33342 on L1210 cells in vitro

et al. 1988

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This study was designed to evaluate the effects of vital dye Hoechst 33342 (HO 33342), at concentrations used to obtain a good DNA histogram resolution, on DNA integrity, cell growth, and cell-cycle phase distribution of Ll210 cells. HO 33342 exposure for 2 h, at 37°C produced DNA single-strand breaks as assessed by the method of alkaline elution. DNA single-strand breaks were concentration dependent (in the range .5-5 pg/ml) and increased significantly when HO 33342 (0.5-1.5 pg/ml) was associated with exposure in a flow cytometer to U.V. laser beam illumination.HO 33342 produced a cytotoxic effect on cell growth even at the concentration of 0.5 pg/ ml-a concentration ten-fold smaller than those required to obtain a good DNA histogram resolution. HO 33342 produced a severe block of the cells in the Gz-M phase of the cell cycle already evident 24 h after stain exposure and continuing up to 144 h after start of recovery. A new polyploid cell population (with a 4 c DNA content) not present in the unstained cells was already evident 24 h after dye exposure. The data shown in the present paper would imply caution in using sorted cells stained with HO 33342 dye for biological, biomedical, and pharmacological studies.Key terms: Flow cytometry, danger in staining Among the DNA probes used in flow cytometry, the bis-benzirnide Hoechst 33342 (HO 33342) dye is of particular interest as it permits vital staining of the DNA structure of cells. HO 33342 was introduced for flow cytometry analysis of living cells by Ardnt-Jovin and Jovin (1); HO 33342 binds to DNA without intercalating, being specific for the A-T sequences (11-15). Upon binding to DNA its quantum efficiency is increased 60-fold and 97% of the fluorescent emission from dyetreated living cells originates from the nucleus (1). Studies with synthetic polynucleotides also showed that not only is a sequence of three A-T pairs necessary for a high dye-binding affinity, but their structural arrangement in a helix is also of importance (18). HO 33342 has been used in many investigations including analysis of DNA content in living cells (1,10,12), sorting of cells with respect to their fluorescence in terms of the DNA content or cell-cycle phase distribution (8,17), and in measures of cell proliferation after drug treatment or irradiation (17J9).Due to its spectral fluorescence characteristics it can be used in conjunction with rhodamine for rapid analysis of cellular DNA and proteins (3, or Despite preliminary studies which suggested that this DNA probe was suitable for vital DNA staining and sorting without affecting cellular integrity (l), it now appears that cellular toxicity and cell-cycle perturbations due to HO 33342 vary considerably from one cell line to another, and that the concentration of this dye required to obtain a good DNA histogram resolution is also cell line dependent (3,6,9,20,22,23,26).Duran and Olive have also shown that concentrations greater than 5 pM of HO 33342 were mutagenic on Chinese hamster V79 lung fibroblast cells, whereas DNA damage, a...

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Section: Discussionmentioning

confidence: 71%

DNA damage, cytotoxic effect and cell‐cycle perturbation of Hoechst 33342 on L1210 cells in vitro

et al. 1988

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“…7,8 Similarly, many other modern chemotherapeutic agents have been isolated from plants to treat cancer in clinical situation successfully. [9][10][11][12][13] Recently, scientists and medical professionals have shown increasing interest in this field as they recognize the true health benefits of herbal medicine, which has been playing an important role in the human healthcare since time immemorial.…”

Section: Introductionmentioning

confidence: 99%

Anticancer activity of Helicia nilagirica bedd in mice transplanted with Dalton’s lymphoma

Jagetia¹

2018

IJCAM

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Cancer afflicts everyone and is a dreaded disease as it does not have definite cure, especially for solid tumors in the advanced stages of maligncies. Modern chemotherapeutic regimens are known to induce adverse toxic side effects in the patients undergoing chemotherapy indicating the need to search newer treatment modalities which are less toxic and would cure cancer. The toxic effects of different extracts of Helicia nilagirica were studied by determining the acute toxicity in normal Swiss albino mice, where mice were injected with different doses of the chloroform, ethanol and aqueous extracts of Helicia nilagirica intraperitoneally. The LD50 was found to be 2 g/kg b. wt. for chloroform and 0.75g/kg b. wt. for aqueous extract, whereas the ethanol extract was nontoxic up to a dose of 2g/kg b. wt. The estimation of anticancer activity of Helicia nilagirica in the Dalton's lymphoma tumor bearing mice showed that administration of 50, 75, 100, 125,150 or 175mg/kg b. wt. aqueous extract of Helicia nilagirica resulted in a dose dependent increase in tumor free survival. The highest survival of 16.6% was observed in the mice receiving 175mg/ kg b. wt. aqueous extract, where tumor free survivors were recorded beyond 120 days with an average survival time of 55 and median survival time of 86 days. The administration of different doses of ethanol extract of Helicia nilagirica also elevated the tumor free survival however only up to 38 days for 175mg/kg b. wt. and no tumor free survivors were observed thereafter. The dose of 175mg/kg. b. wt. aqueous extract was chosen for further investigation as maximum tumor free survivors were observed for this dose. The analysis of micronuclei frequency showed a time dependent elevation up to 24h and a decline thereafter. The apoptosis index also increased with assay time up to 36h post treatment. The liver and kidney function tests did not show any significant alteration indicating that it did not exert toxic effect on these two vital organs. Our study demonstrates that Helicia nilagirica is non-toxic and increase the tumor free survival beyond 120 days by inducing DNA damage, apoptosis and necrosis in the tumor cells.Citation: Jagetia GC, Zoremsiami J. Anticancer activity of Helicia nilagirica bedd in mice transplanted with Dalton's lymphoma. Int J Complement Alt Med.Citation: Jagetia GC, Zoremsiami J. Anticancer activity of Helicia nilagirica bedd in mice transplanted with Dalton's lymphoma. Int J Complement Alt Med.

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“…Vinca alkaloids are active against many solid tumors, and hematological malignancies can be effectively treated using them [8][9][10]. Epipodophyllotoxins have been reported to be active in several rodent and human neoplasms, including Hodgkin's disease, small cell lung carcinoma, testicular tumors, diffuse histiocytic lymphoma, testicular cancer, and certain lymphomas [11][12][13]. Taxol is an antimitotic agent and cells resistant to vinca alkaloids because of mutations in tubulin remain sensitive to taxol.…”

Section: Introductionmentioning

confidence: 99%

Short - Term Loosen Up Meditation Induced EEG and Autonomic Response in Healthy Japanese Students

Iwakuma¹,

Nakayama²,

Oshita³

2016

J Alt Med Res

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Back ground: Cancer has emerged as the second largest killer disease in the world and mortality rates for solid tumors remain unchanged. Therefore there is need to screen, new remedies to treat cancer. Materials and methods:The antineoplastic activity of different doses of hydroalcoholic extract of Alstonia scholaris (L.) R. Br. (ASE) was studied in Swiss albino mice, transplanted with Ehrlich Ascites Carcinoma cells (EAC) in its peritoneal cavity. The effect of ASE injected at different stages of tumor development was evaluated by studying tumor free survivors. In another study EAC cells were treated with Echitamine Chloride (ECL) in vitro and transplanted back into mice and tumor free survivors determined. The glutathione and lipid peroxidation were also measured after ASE treatment. Results:The daily administration of ASE in tumor bearing mice caused a dose-dependent remission of tumor, and highest regression was observed at 480 mg/kg b. wt. this dose showed some toxic effects and next lower dose i.e. 420 mg/kg was considered as the suitable dose, where 33.33% of the animals survived up to 120 days post-tumor-cell inoculation, as against no survivors in the saline treated control and positive cyclophosphamide treated groups. The ASE caused a dosedependent elevation in the median survival time (MST) and average survival time (AST), causing a corresponding increase in the median life span (IMLS) and average life span (IALS) of experimental animals. Treatment of EAC mice with 420 mg/kg ASE also retarded tumor development even during the late stages of tumor development, while cyclophosphamide was completely ineffective. The in vitro treatment of EAC cells with ASE or ECL and their transplantation in mice showed that ASE treatment was more potent than ECL treatment as evident by increased MST and AST in the former when compared with the latter. Assay of glutathione and lipid peroxidation after 240, 420 mg/kg ASE or 25 mg/kg cyclophosphamide led to a decline in glutathione contents and increase in lipid peroxidation. Conclusions:Our study shows that ASE treatment kills the tumor cells effectively increasing long-term disease free survival of experimental mice, which may be due to reduced glutathione and increased lipid peroxidation.

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Flow-cytometric analysis of DNA distribution after VP16-213 treatment of Lewis lung carcinoma

Cited by 17 publications

References 11 publications

DNA damage, cytotoxic effect and cell‐cycle perturbation of Hoechst 33342 on L1210 cells in vitro

DNA damage, cytotoxic effect and cell‐cycle perturbation of Hoechst 33342 on L1210 cells in vitro

Anticancer activity of Helicia nilagirica bedd in mice transplanted with Dalton’s lymphoma

Short - Term Loosen Up Meditation Induced EEG and Autonomic Response in Healthy Japanese Students

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