Inhibition of Human Breast Carcinoma Growth by a Soluble Recombinant Human CD40 Ligand
Akio HiranoDan L. LongoDennis D. TaubDouglas K. FerrisLawrence S. YoungAristides G. EliopoulosAngelo AgathanggelouNicky CullenJ. C. MacartneyWilliam C. FanslowWilliam J. Murphy
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Breast carcinoma
In recent years, the potential of nanobubbles (NBs) for biological activation has been actively investigated. In this study, we investigated the proliferative effects of nitrogen NBs (N-NBs) on fibroblast cells using cell assays with image analysis and flow cytometry. A high concentration of N-NBs (more than 4 × 108 NBs/mL) was generated in Dulbecco's modified Eagle's medium (DMEM) using a gas-liquid mixing method. In image analysis, the cells were counted and compared, which showed an 11% increase in cell number in the culture medium with N-NBs. However, in two further cell cytometry analyses, the effect of nanobubbles on cell division was found to be insignificant (approximately 2%); as there is insufficient evidence that N-NB is involved in cell division mechanism, further studies are needed to determine whether NB affects other cellular mechanisms such as apoptosis. This study presents the first successful attempt of directly generating and quantifying N-NBs in a culture medium for cell culture. The findings suggest that the N-NBs in the culture medium can potentially facilitate cell proliferation.The online version contains supplementary material available at 10.1007/s13534-022-00242-y.
Cytometry
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Several human and murine tumor cell lines were evaluated in an in vitro cytotoxicity assay as prescreens for fermentation extracts and pure materials subsequently tested in vivo against P388 leukemia or B16 melanoma. Each material, regardless of its in vitro cytotoxicity, was evaluated in vivo. At the criteria levels of in vitro positivity and in vivo activity invoked, a highly significant relationship between these two endpoints was demonstrated for each cell line. When cell lines were compared, most of them performed in a similar manner, with HCT-116 human colon carcinoma cells providing a modest advantage predicting for P388 activity in some comparisons. Using the data from any two cell lines in concert did not improve the acuity of the prescreen beyond that associated with the better cell lines used singularly and only a minority of active materials was predicted for uniquely. Overall, the in vitro cytotoxicity assay provided a useful prescreen for selecting P388 and B16 in vivo active materials.
In vitro toxicology
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Abstract Enhancement of the anti‐proliferative effect of human interferon (HuIFN) preparations (α, β and γ) by dipyridamole was detected in a human malignant melanoma cell line, MMICB, which we originally established. Cell growth was inhibited by HulFN alone, but a marked increase in inhibition was noted in vitro and in vivo when dipyrydamole was added. Cellular DNA synthesis, as dletermined by 3 H‐deoxythymidine incorporation into the acid‐insoluble cellular fraction, was more inhibited by combined treatment than by any of the agents used alone. Two other melanoma cell lines that we established, MM‐2CB and MM‐3CB, also exhibited sensitivity to combined treatment both in vitro and in vivo. Furthermore, the HMV‐I and SEKI melanoma cell lines were susceptible to the combination. Even non‐cytotoxic concentrations of dipyridamole could enhance the effect of HulFN on MM‐ICB, MM‐2CB, and SEKI cells.
Dipyridamole
Thymidine
Interferon alfa
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Immunosuppression
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To investigate the inhibitory effect of the extractive of Ammopiptanthus mongolicus(JA1)on hepatocarcinoma cells and its possible mechanisms,the hepatocellular carcinoma cell line of mice(H22)was used as target cell.The inhibitory effects of JA1 on H22 cells were assayed by MTT in vitro and the antitumor effect of JA1 was also observed in H22-bearing mice by flow cytometry assays,light and transmission electronic microscopy.The results revealed that JA1 had significant anti-proliferative effects on H22 cells with the dependence of time and dosage.In addition,JA1 not only significantly inhibited the proliferation of H22 cells in vitro but also inhibited the growth of H22 hepatocarcinoma in vivo.Flow cytometric analysis indicated that JA1 could induce apoptosis on H22 cells in H22-bearing mice and the G1 phase cell percentage increased and the S phase cell percentage decreased.Morphological change of H22 cells undergoing apoptosis were also detected by light and electron microscopy.These results suggest that JA1 has better inhibitory effects on H22in vitro and vivo and the mechanism maybe related to its apoptosis inducing activity.
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Objective To study the effects of saikosaponins-d(SSd) on proliferation and cell cycle in the human hepatocellular carcinoma cell line,HepG2 cell line.Methods HepG2 cells were cultured and treated with SSd.The cell growth rate was measured by monotetrazdium(MTT) assay and the cell cycle was determined by the flow cytometry(FCM) analysis.Results The SSd obviously inhibited the proliferation of HepG2 cell line in a concentration and time dependent manner,the maximum response of the cells to SSd was at the concentration of 10mg/L and the treatment time-point of 48h.Compared with the control,the cells treated with 10mg/L SSd for 48h significantly reduce cell numbers in S phase (P0.01).Conclusion The SSd could inhibit the proliferation of HepG2 cell line and arrest its cell cycle in S phase.
Hepatic carcinoma
Cell counting
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Relationship between tumor cell growth and cell cycle and influence of antitumor drugs on the cell cycle regulation were investigated by the use of murine EL-4 tumor. Cell growth curve of EL-4 cells was well related to the cell cycle pattern analyzed by two color flow cytometry both in vivo and in vitro. The ratio of S phase rapidly increased in early log phase and decreased from late log to plateau phases. The ratio of G0/G1 phase showed a reciprocal change. Minimum effective doses of 5-fluorouracil (5-FU) and CDDP on EL-4 growth in vitro were 1 x 10(-6) M and 0.5 micrograms/ml, respectively. At such doses, 5-FU showed an accumulation in early S phase and CDDP showed a partial synchronization in S phase and subsequent accumulation in late S and G2 + M phase. In case of in vivo administration with 5-FU (20 mg/kg/day) ratio of S phase was higher than in untreated control mice at day 2-3 but decreased rapidly thereafter. Mice administered with CDDP (5 mg/kg/day) showed a decrease in S phase from day 2 and completely rejected the tumors by day 5. From these results, each phase of cell cycle was influenced by the cell growth characteristics and the cell cycle pattern was dynamically changed according to the mode of action of antitumor drugs. Moreover, in vivo effects of these drugs can be evaluated adequately by the analysis of the cell cycle.
S phase
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AIM To study the effects of crude extracts of Aspongopus chinensis Dallas on cell proliferation and cell cycle of SGC-7901 and HepG2 cell lines.METHODS The crude extracts were extracted by chloroform infusion.The antiproliferative effects of crude extracts were assessed by MTT assays.The cell cycle was measured by flow cytometry.RESULTS The crude extracts could inhibit the proliferation of SGC-7901 and HepG2 cell lines in a dose-dependent manner.SGC-7901 and HepG2 cell lines were sensitive to crude extracts with an IC50 of 1 193.52 and 964.34 μg/mL,respectively.From the result of flow cytometry analysis,crude extracts,as compared with the untreated cells,caused significant reduction in cell ratio at the S and G2/M phases,and an increase in cell ratio at the G0/G1 phases.CONCLUSION The crude extracts could inhibit proliferation of both cancer cell in vitro,and especially arrest human liver cancer cells at the G0/G1 phase of the cell cycle for HepG2.
MTT assay
G1 phase
IC50
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Objective To study the inhibition effect of PC on human cholangiocarcinoma in vivo and vitro.Methods Cells were cultivated in vitro,and then were divided into positive control group,negative control group and PC group randomly after cultured for 24 h.The cytostatic effect of PC on human cholangiocarcinoma w as evaluated by MTT assay.Flow cytometry w as utilized for measuring the apoptosis and cell cycle.The nude mice w ith QBC939 cell xenografts w ere divided into 5 groups randomly: negative control group,positive control group,PC groups(three dosage groups),and the mice w ere treated w ith drugs via intraperitoneal injection daily.The tumor size w ere measured every three days.After 12 d of treatment,the tumor was weighed,and the inhibitory rate w as calculated.Results PC inhibited cell grow th in a concentration-and-time dependent manner.PC blocked cell cycle in phase S,and elevated the apoptosis rate,PC remarkably inhibited the grow th of QBC939 cell xenografts.Conclusion PC can inhibit the grow th of SHG-44 in vitro and in vivo,and induce the apoptosis of QBC939.
MTT assay
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To study the regulation role of tumor suppressor NECL1 on the proliferation of glioma cell line.We detected the expression level of NECL1 in human normal brain tissue and glioma cell lines using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot. T98G cell line in which NECL1 was silent and whose transfection efficiency was relatively high as target cell was chosen, and the effect of NECL1 on the proliferation of T98G cell line in vitro was detected by using cell growth curve, flow cytometry, and Hoechst staining.NECL1 was abrogated or markedly reduced in 6 glioma cell lines. When NECL1 was overexpressed in T98G cell line, the cell growth rate obviously decreased and the number of apoptotic cells remarkably increased when compared with the control group.NECL1 may inhibit the proliferation of T98G cells by inducing its apoptosis.
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