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Aim To compare high-content cell imaging system and other methods in detecting cell proliferation, including the traditional thymidine ( 3H-TdR) incorporation method, methyl thiazolyl tetrazolium ( MTT ) method and cell counting kit-8 (CCK-8) method. Methods The fibroblast-like synovial cells (FLS) were used as the study object to observe the sensitivity and stability of FLS proliferation in different methods by using the usual proliferative stimulant tumor necrosis factor-α ( TNF-α) and the known proliferation inhibitor methotrexate at different concentrations. Results The 3H-TdR method and high-content cell imaging could detect a significant inhibitory effect of 1 nmol ·L-1 MTX on FLS cell proliferation, MTT assay and CCK-8 method could detect the significant inhibitory effect of 10 nmol· L-1 MTX on FLS cell proliferation. 3H-TdR method was found to have a large degree of discretization in the data set, with a standard deviation of 32.61% ~61.36% , and the MTT method was 11.9% ~ 17.8% , the CCK-8 method was 17.15% ~32.88% , and the high-content cell imaging system method was 12.66% ~26.54%. Conclusion The method of high-content cell imaging system is more accurate and stable for detecting cell proliferation.
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Objective To compare the incorporation method of 3H-TdR and 125Ⅰ-UdR on determining the proliferation effect of lymphocytes. Methods The proliferation effects of lymphocyte and Daudi lymphoma cells were estimated by 3H-TdR and 125Ⅰ-UdR incorporation. Results The incorporating fraction of 3H-TdR and 125Ⅰ-UdR into lymphocyte was 20.95% ± 1.06% and 1.00% ±0.04%,respectively, and the incorporating fraction for the lymphoma cells was 29. 94% ± 4. 10% and 6. 02% ±0. 73% ,respectively. The incorporation fractions of 3H-TdR into lymphocyte and lymphoma cells were much higher than those of 125Ⅰ-UdR, but the incorporating fractions of 3H-TdR or 125Ⅰ-UdR into the lymphoma cells were much higher than those of lymphocytes. Conclusions For lymphocytes, 125Ⅰ-UdR cannot substitute 3H-TdR as a tracer agent. But for lymphoma cells, whether 125Ⅰ-UdR could be replace 3H-TdR or not needs further research.
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Aim To estimate the anti-uterine cervix cancer effects of the extracts from Lysimachia clethroides Duby in vivo and in vitro. Methods Inhibitory effect on growth was observed by MTT, 3H-TdR and colony-forming units assay. Apoptosis was detected by Hoechst fluorescent staining analysis. The effect on cell migration and morphology was observed by scratch test; Detecting effects of ZE4 on transplant tumor (U14) in mice through observing the tumor weight and organ index. Results ZE4 could inhibit the growth and migration of HeLa cell significantly and induce cell apoptosis. Its half inhibitory concentration (IC50) at 48h was 40.56 mg?L-1 by 3H-TdR assay. The inhibition rate of ZE4 against U14 was 49.9% at the dose of 400 mg?kg-1. Conclusions ZE4 could inhibit the growth of uterine cervix cancer in vitro and in vivo.
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Purpose:To study the effect of sodium selenate on the viability and proliferation of PC-3 cell, a kind of human prostate cancer cell. Methods:Sodium selenate was administered to PC-3 cells, and MTT assay and ~(3)H-TdR adulterated assay were used to estimate the viability and proliferation of cell. Results:① When cells were treated with Sodium selenate for 24 h, the optical density (A) of middle-dose group decreased significantly, and the A of the high-dose group decreased dramatically. When cells were treated for 24 h, the A of the low-dose group was significantly lower than that of the control group, while the A of the middle-dose and high-dose groups was much lower than control.② When cells were treated for 24 h, the proliferation of middle-dose group decreased, and that of high-dose group decreased markedly. Conclusions:Sodium selenate can inhibit the viability and proliferation of PC-3 cells, and these actions occur in a dose-dependant manner.
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OBJECTIVE:To study the effect of ligustrazine injection(LTZ)on DNA synthesis of vascular endothelial cell.METHODS:In vitro cultured cell line of human umbilical vein endothelial cells,ECV304,were adopted to measure the ef?fects of LTZ on DNA synthesis by 3 H-TdR incorporation.RESULTS:LTZ could inhibit DNA synthesis of ECV304in a dose-dependent manner.CONCLUSION:The mechanism of anti-angiogenesis of LTS might be associated with inhibiting DNA synthesis of vascular endothelial cell.
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PURPOSE. To evaluate the effects of interfe-ron-gamma (IFN-?) and interferon-alpha (IFN-?) on human retinal pigment epithelial (RPE) ceils in vitro. METHODS: Cultures of human RPE cells were incubated with either of the IFNs (10IU/ml～10~4 IU/ml) for 3～5 days, and proliferation rates of the cells were assayed by [~3H]-thymidine incorporation and liquid scintillation techniques. RESULTS: There was a slight inhibition on human RPE cells with dosages from 10IU/ml to 10~4IU/ml. The inhibition rates of IFN-? of 10~3IU/ml, 10~4IU/ml were 8%, 15%, 26%, and of IFN-? were 7%, 9%, and 16%, respectively. On the 5th day there was a slight decrease in the inhibition rate than on the 3rd day. CONCLUSION: IFN-? and IFN-? exerted a slight inhibition on human RPE cells lasting 3 days.
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A rapid, sensitive, reliable and simple microdetermination technology for anti-cancer activity, incorporation technology of ~3H-TdR, was developed to seek the process for anti-cancer preparations of compound Chinese medicines. In such a way, more rational process could be drafted during the short time. The preparation made by this process was found to possess good curative effect, no obvious toxicity and no side-effect in the compreshensive evaluation of clinic and a total 14 biology indexes, The application of some rapid and sensitive medical-determination technology in studies of preparation process, quality standards and stability could solve some problems induced by indefinite active components, make research way broad, and raise research level.
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The activity of colony stimulating factor ( CSF) in the conditioned medium ( CM ) was studied with combination method of 3H-TdR incorporation assay and agar colony assay. These two assays were demonstrated to be replaceable each other. The mice were administered with nontoxic monophosphoryl lipid A(MPLA) which was derived from a Re mutant of Salmonnella Minnesota Re595. The results showed a significant elevation of CSF in the serum and reaching the top at 12th h and returning to normal by 24th h. There is a significant dose-resoonse relationship. The CSF was induced with accompany of formation of colony inhibiting factor (GIF), some of which were heat sensitive factors. It is suggested that the MPLA may be a potent CSF-inducer.
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The influences of varied concentrations of AG on the 8H-TdR(3H-Thymidine ) incorporation inhibition rate and the release of PGI2 and TXA2 were studied in cultured human and rat lymphocytes . The results indicated that the AG increased the 3H-TdR incorporation inhibition rate on the concentrations of 1.5? 10-7mol/L and 3?10-7mol/L. Using the RIA method, authors find that AG on the concentrations of 4 ? 10-7mol/L, 4 ? 10-3mol/L and 4 ? 10-11mo/L can significantly decrease the cultured lymphocytes TXA2 release while has no clear influences on PGI2 release
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AIM To study the effect of A 1015 on D galactosamine induced mice hepatotoxicity. METHODS The effect of three doses of A 1015 on DNA biosynthesis of D galactosamine induced mice hepatotoxicity were tested. The time curve of A 1015 on DNA synthesis of D GL induced mice hepatotoxicity was also observed. RESULTS The experiments demonstrated that 2 5 mg?kg -1 is the optimum dose for A 1015 to show liver protective activity. The results suggested that HGF can not only offset the hepatotoxicity induced by D GL, but also promote the DNA synthesis of mice liver cells. And the highest DPM value for HGF group is higher than that of A 1015 group. Thus both HGF and A 1015 showed anti hepatotoxic activity and promoting activity on mice liver cell DNA synthesis. CONCLUSION A 1015 may reduce liver necrosis induced by D galactosamine and promote the DNA synthesis of mice liver cells.
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In this study, we observed the efTects of systemic poisoning by sulphur mustard (SM) on DNA biosynthesis of internal organs and protective action of antidotes in mice.The results showed that the systemic poisoning by SM produced a strong depression of [3H]-TdR incorporation into DNA biosynthesis, which was characterized by rapidity, severity and rapid recovery.It is suggested that mammal an organism has a marked physiological compensation, regeneration and repair activity for DNA damage.Antidote sodium thiosufate alone or in combination with unithiol (DMPS) has satisfactory protective effects on depression of DNA biosynthesis in mice poisoned by SM.
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We studied the changes of colony-stimu lating factor (CSF) level, the type of CSF after intraperi-toncal injection of estriol sodium succinate (E3?SNa) and the effect of E3?SNa itself on the proliferation of bone marrow precursors with the use of both agar colony assay and [3H]-TdR incorporation assay. Our experiment shows that CSF was at high level 6 h after E3?SNa administration and reached a peak by 24 h, and that elevation of CSF in the serum was maintained for at least 30 d. Using 0.14-28 mg of E3?SNa there was a dose-dependent increase in serum CSF when tested 24 h after E3?SNa treatment. The main type of CSF was GM-CSF. There was no direct stimulation of E32?SNa itself on the proliferation of mouse bone marrow precursors. So we think that the effect of estrogens on the hematopoiesis may, at least in part, be mediated by altered CSF activity.
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5srRNA was isolated and purified from the reticulocytes of rabbit. It labeled with ~(125)I, then incubated with mouse myeloma cells (SP2/0). By autoradiography it was observed that ~(125)I-SsrRNA could pass into the nuclei of the cells. In a separate experiment, it showed that the incorporation rate of ~3H-TdR into nuclei of SP2/0 after incubation with 5srRNA was decreased as compared with that f control group, hence the result indicates that 5srRNA inhibits DNA synthesis of the SP2/0 cells and it seems to play a role in the regulation of gene expression through its hybridization with DNA sequences of the SP2/0 cells. Thus it is likely that 5srRNA might act as "erythroid denucleation factor".
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The changes of the growth characteristics and surface ultrastructure during longterm passages of Wg3h cells that have been transfected with PSV_2-neo (Wg3h-neo) were studied. The results showed that the growth and DNA synthesis rate were evidently higher in the transfected cells than in the parental wg3h cells. The saturation of the transfected cell growth density was also increased, but neither the parental and transfected cells formed cell colony in soft agar medium nor tumor growth in nude mice. There was no significant differences in morphology between the two cell types under light microscopy, however much more microvilli were seen on the transfected cell surface under scanning electron microscopy. Southern blot hybridization analysis indicated that the PSV_2-neo plasmids were integrated into the genome of the target cells. Our results demonstrated that the transfected cells remained as nonmalignant cells although some changes of their growth characteristics and surface ultrastructure appeared.