Effect of Arsenic Trioxide Combined with Dihydroartemisinin on THP-1 Cells Proliferation and Apoptosis / 中国实验方剂学杂志

Objective:To investigate the effects of arsenic trioxide combined with dihydroartemisinin on proliferation, cell cycle, and apoptosis of THP-1 cells, and explore the mechanism. Method:The thiazolyl blue （MTT） method was applied to detect the effect of different concentrations of arsenic trioxide, dihydroartemisinin and arsenic trioxide combined with dihydroartemisinin on the proliferation of THP-1 cells. Annexin V/propidium iodide（PI）assay was used to detect the change of THP-1 cell cycle and apoptosis．Western blot was performed to assess the expression of cysteine protease-3（Caspase-3）, cleaved Caspase-3, B-lymphocytoma-2（Bcl-2） and Bcl-2 associated X protein （Bax）. The changes of cell morphology were observed under high intension microscope. Result:Compared with blank group, arsenic trioxide and dihydroartemisinin both exhibited obvious antiproliferative effect on the human acute monocytic leukemia cell line THP-1 in time-dose dependence （P<0.01）. After 48 h, compared with the same dose of arsenic trioxide or that of dihydroartemisinin alone, the inhibition effect of 1 µmol·L-1 arsenic trioxide combined with 2 µmol·L-1 dihydroartemisinin on proliferation of THP-1 cells was significantly stronger （P<0.01）. Compared with the control group, arsenic trioxide combined with dihydroartemisinin significantly arrested the cells in G1 phase （P<0.01）, induced the downregulation of Caspase-3 and Bcl-2 （P<0.01） and upregulation of cleaved Caspase-3 significantly（P<0.05）. Conclusion:Arsenic trioxide combined with dihydroartemisinin can significantly inhibit the proliferation and induce apoptosis of THP-1 cells. The possible mechanism may be related to arrest the cells in G1 phase, reduce the expression of Caspase-3 and Bcl-2, increase the expression of cleaved Caspase-3.

Preparation and in vitro evaluation of redox-responsive silica-loaded arsenic trioxide nano-drug delivery system / 中草药

Objective: The redox-responsive drug delivery system of MSN-SS-PEG@As2O3 was constructed based on mesoporous silica nanoparticle (MSN), which was modified by both redox-sensitive disulfide bonds and non-toxic, non-immunogenic polyglycols (PEG), and loaded the arsenic trioxide (As2O3) by electrostatic adsorption and evaluated in vitro. Methods: Silica was synthesized by coprecipitation method. The redox-responsive carrier (MSN-SS-PEG) was synthesized on the basis of silica, (3-mercaptopropyl) trimethoxysilane, 2-(2-pyridyldithio) ethylamine hydrochloride and methoxy terminated PEG. The particle size and Zeta potential of MSN-SS-PEG were measured by Malvern particle size analyzer; The structure of the carrier was verified by infrared spectroscopy; The morphology and physical and chemical properties of the carrier were investigated by transmission electron microscopy (TEM) and small angle powder diffraction; The drug loading efficiency of MSN-SS-NH2@As2O3 and MSN-SS-PEG@As2O3 were investigated by inductively coupled plasma emission spectrum (ICP). The drug loading was further verified by thermogravimetry (TGA). In vitro release characteristics of the drug delivery system under different pH conditions were investigated by dialysis bag method. MTT assay was used to investigate the toxicity of carrier and delivery system to human normal hepatocytes (L02) or human hepatocarcinoma (HepG2) cells. Results: The potential of MSN, MSN-SS-NH2, MSN-SS-PEG was (-13.40 ± 0.87), (31.63 ± 0.90), (27.70 ± 5.60) mV, respectively. The final potential of modified carrier was positive. The particle size of MSN-SS-PEG was (159.60 ± 3.10) nm. The results of TEM showed that MSN, MSN-SS-NH2 and MSN-SS-PEG were all round or quasi round; The drug loading of MSN-SS-PEG@As2O3 was 4.38%, which measured by ICP; The release in vitro showed that MSN-SS-PEG@As2O3 was redox sensitive response. Compared with L02 cells, HepG2 cells were more sensitive to the toxicity of the carrier, and with the increase of the carrier concentration, the cell survival rate of MSN-SS-PEG was higher than that of MSN-SS-NH2, suggesting that PEG modification can further reduce the cytotoxicity of the carrier and improve the biocompatibility of the carrier. In addition, MTT results showed that the inhibitory effect of MSN-SS-PEG@As2O3 on HepG2 cell was significantly higher than that of other groups. Conclusion: The carrier prepared in this paper had a round and uniform particle size. The modified silica can release under the special microenvironment of the tumor and increase the accumulation of the drug in the tumor site. The delivery system has a good application in tumor therapy as a tumor micro-environment responsive carrier.

Quality Control of Realgar in Compound Huanglian Ointment / 中国中医药信息杂志

Objective To control the quality of realgar in compound Huanglian Ointment.Methods As2O3which is the toxic component of realgar was carried limit test by Gutzeit's test; Potassium sulfate, ammonium sulfate and sulfuric acid were used to digest, and then titration method was used to determine the content of As2S2in compound Huanglian Ointment. Results The content of the soluble As in compound Huanglian Ointment was no higher than 15.6 μg/g. The content of As2S2in compound Huanglian Ointment was no less than 2.21 mg/g. Conclusion The method is simple and easy,which can be applied as the quantity control method of compound Huanglian Ointment.

Influence of As2O3 combined with ginsenosides Rg3 on inhibition of lung cancer NCI-H1299 cells and on subsistence of nude mice bearing hepatoma

OBJECTIVES@#To study the effect of arsenic trioxide (As2O3) combined with ginsenosides Rg3 on inhibiting the NCI-H1299 lung cancer cells and subsistence in nude mice bearing hepatoma.@*METHODS@#MTT method was used to measure the inhibition effect of As2O3 combined Rg3 on NCI-H1299 cells, and the proliferation inhibiting effect was observed via establishing the transplanted tumor model in vitro. A total of 40 tumor-bearing nude mice were randomly divided into normal saline group, As2O3, Rg3 and As2O3+Rg3 group. Transplantation tumor model of lung cancer in nude mice was constructed, followed by injection of certain concentrations of normal saline, As2O3, ginseng saponin Rg3 and As2O3+Rg3 every day. The survival duration and the tumors size of the mice were recorded and the Kaplan-Meier curve was made; microscopic observation of apoptosis of tumor cells in vivo was done using TUNEL staining.@*RESULTS@#After 72 h of injection, inhibition rate of tumor cell in normal saline group, As2O3 group, Rg3 group and As2O3+Rg3 group was (5.66±0.31)%, (65.58±4.75)%, (44.69±3.32)% and (82.67±5.43)%, respectively. Inhibition rate of tumor cell in As2O3 group, Rg3 group and As2O3+Rg3 group was significantly higher than that of normal saline group (P<0.01); inhibition rate of tumor cells of As2O3+Rg3 group was significantly higher than that of the two groups given As2O3 or Rg3 alone (P<0.01). The tumor volume of As2O3 group, Rg3 group and As2O3+Rg3 group shrank to (65.38±3.25)%, (77.68±3.43)% and (42.65±3.55)% of the original, tumor volume of saline group was 1.21 times of the original size (P<0.01); Median survival of saline group, Rg3 group, As2O3 group were significantly shorter than that of As2O3+Rg3 group (P<0.01); co-ordinated intervention ability of As2O3+Rg3 on NCI-H1299 cell was significantly higher than that of As2O3 or Rg3, separately.@*CONCLUSIONS@#As2O3 combined with Rg3 can significantly inhibit proliferation of NCI-H1299 cells in lung cancer, prolong survival of tumor-bearing nude mice, and promote tumor cell apoptosis, and have significant effect on lung cancer treatment.

Inhibitory effect of humanized anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles conjugate on growth of human hepatocellular carcinoma: in vitro and in vivo studies

OBJECTIVE@#To investigate the inhibitory effect of humanized anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles conjugate on growth of human hepatocellular carcinoma both in vitro and in vivo, which may be a potential agents with sensitivity and targeting ability for human hepatocellular cancer.@*METHODS@#Humanized anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles conjugate was previously constructed using ribosome display technology and antibody conjugate technology. In this combined in vitro and in vivo study, the inhibitory effects of anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles conjugate on tumor growth, invasion, and metastasis was observed with human liver carcinoma cell line Bel7402 and normal cell L02 by MTT assay, Tanswell assay, Hochest33258 staining, and DNA ladder analysis. The anticancer activity and distribution of anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles was then verified in a mouse model of Bel7402 xenografts.@*RESULTS@#Anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles significantly inhibited the proliferation of Bel7402 in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay while had almost no effects on L02 cells. And the apoptosis inducing effects were proved by Hochest33258 staining and DNA ladder analysis. Transwell assay found that the drug also inhibited the metastasis ability of tumor cells. Furthermore, anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles significantly delayed the growth of Bel7402 xenografts after administration (92.9%), followed by As2O3-stealth nanoparticles, anti-VEGFR-2 ScFv, and As2O3 (61.4%, 58.8%, 20.5%, P<0.05). The concentration of As2O3 in anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles group was more selectively.@*CONCLUSIONS@#Anti-VEGFR-2 ScFv-As2O3-stealth nanoparticles is a potent and selective anti-hepatocellular carcinoma agent which could inhibit the growth of liver cancer as a targeting agent both in vitro and in vivo and also significantly inhibit angiogenesis.

ETME, a novel β-elemene derivative, synergizes with arsenic trioxide in inducing apoptosis and cell cycle arrest in hepatocarcinoma cells via a p53-dependent pathway

Arsenic trioxide (ATO) has been identified as an effective treatment for acute promyelocytic leukemia (APL) but is much less effective against solid tumors such as hepatocellular carcinoma (HCC). In the search for ways to enhance its therapeutic efficacy against solid tumors, we have examined its use in combination with a novel derivative of β-elemene, N-(β-elemene-13-yl)tryptophan methyl ester (ETME). Here we report the effects of the combination on cell viability, apoptosis, the cell cycle and mitochondria membrane potential (MMP) in HCC SMMC-7721 cells. We found that the two compounds acted synergistically to enhance antiproliferative activity and apoptosis. The combination also decreased the MMP, down-regulated Bcl-2 and pro-proteins of the caspase family, and up-regulated Bax and BID, all of which were reversed by the p53 inhibitor, pifithrin-α. In addition, the combination induced cell cycle arrest at the G2/M phase and reduced tumor volume and weight in an xenograft model of nude mice. Overall, the results suggest that ETME in combination with ATO may be useful in the treatment of HCC patients particularly those unresponsive to ATO alone.

Effect of As2 O3 and paclitaxel on the proliferation of A549 cell and ectopic tumor growth of nude mice / 中国生化药物杂志

Objective To evaluate the effect of As2O3 combined with paclitaxel(PTX)on the treatment of lung cancer.Methods The anti-proliferation efficiency of As2 O3 combined with PTX was evaluated by MTT assay.Tumor spheroids were used to evaluate anti-tumor ability of As2 O3 combined with PTX.Transmission electron microscope (TEM)were used to observe the apoptosis morphous.A549 cell were xenografted in mice to establish the animal model,and the nude mices were devided into four groups,saline group,As2 O3 group,PTX group and As2 O3 +PTX group.The animal model were used to evaluate the effect of anti-tumor.The tumor size of every group were measured.HE was used to observe the apoptosis of cancer cells. Results The cell inhibition rate of A549 cell were(3.35 ±0.21)%,(47.55 ±2.25)%,(64.64 ±3.35)%and(84.58 ±3.76)%after treatment with saline,As2O3,PTX and As2O3combined with PTX after 48h respectively(P<0.01).The early apoptosis rate of cancer cells were 0.26%,9.7%, 17.8% and 42.5% for saline group,As2 O3 group,PTX and As2 O3 +PTX group respectively(P<0.01 ).The final tumor spheroid volumes in saline group increased 1.36 times after 7 days.The final tumor spheroid volumes reduced to(77.35 ±2.31)%,(61.68 ±2.44)% and(44.85 ±3.34)% in As2O3,PTX and As2O3 combined with PTX group respectively(P<0.01).The inhibition of lung cancer in vitro demonstrated the inhibition rate of tumor growth compared with saline group were(22.4 ±4.5)%,(39.5 ±6.2)% and(69.5 ±7.3)% for As2O3,PTX and As2O3 +PTX,respectively(P<0.01 ).Conclusion As2 O3 combined with PTX can effectively inhibit the proliferation of A549 cells and ectopic tumor growth in nude mice and it may be a potentially effective treatment for lung cancer.

Role and mechanism of autophagy in the arsenic trioxide-induced death of Burkitt lymphoma Raji cells / 中国药理学通报

Aim To investigate the role of autophagy and its mechanism in Raji cell death induced by arse-nic trioxide. Methods Transmission electron micros-copy ( SEM) and MDC fluorescence staining were used to observe autophagy. MTT colorimetry was employed to assay the cellular proliferating activity. Cell apopto-sis and cell cycle analysis were performed using FITC-Annexin-V/PI double staining and flow cytometry ( FCM) . The expressions of LC3 and the conversion of LC3-I to LC3-II were measured by western bloting. The expression of bcl-2 mRNA and p53 mRNA were detected by reverse transcription-polymerase chain re-action ( RT-PCR ) . Results Arsenic trioxide could obviously inhibit the proliferation of Raji cells, arrest the cells at G2/M phase and induce apoptosis. Mean-while, arsenic trioxide markedly inhibited the expres-sion of bcl-2 mRNA and enhanced the expression of p53 mRNA in Raji cells. Arsenic trioxide also induced autophagy synchronously which paralleled with the in-duction of apoptosis in Raji cells, and 3-MA, an auto-phagy inhibitor, was able to reverse the arsenic triox-ide-activated autophagic activity, up-regulate bcl-2, down-regulated p53 expression and suppress the lethal effect of arsenic trioxide on Raji cells to reduce their sensitivity to arsenic trioxide. In contrast, the Rapamy-cin, an autophagy inducer, possessed the completely opposite effects on Raji cells compared with 3-MA. Conclusions The apoptosis and autophagic cell death are coexistent in arsenic trioxide-triggered death of Raji lymphoma cells, and Bcl-2 and p53 may play a key regulating role in this process.

In vitro synergistic effect of arsenic trioxide with conventional or new drugs on the proliferation of cutaneous T cell lymphoma cells Hut-78 and Hut-102 / 中国肿瘤临床

Objective:To investigate the in vitro effect of arsenic trioxide (As2O3) alone and in combination with dexamethasone (DXM), etoposide (VP-16), methotrexate (MTX), bortezomib (BTZ), and suberoylanilide hydroxamic acid (SAHA) on the growth of human cutaneous T cell lymphoma (CTCL) cells Hut-78 and Hut-102. Methods:Hut-78 and Hut-102 cells were cultured with different concentrations of As2O3, DXM, VP-16, MTX, BTZ, and SAHA alone and As2O3 in combination with DXM, VP-16, MTX, BTZ, or SAHA for 48 h. The effects of the different samples on Hut-78 and Hut-102 cell proliferation were determined by MTT assay. Analyses using CalcuSyn software were performed to determine whether the combination of As2O3 with DXM, VP-16, MTX, BTZ, or SAHA in-duced synergistic cytoxicity. Results:As2O3, DXM, VP-16, MTX, BTZ, and SAHA alone significantly inhibited the growth of Hut-78 and Hut-102 cells in a dose-dependent manner, with a 50%inhibiting concentration of 5μmol/L, 500μg/mL, 2.5μg/mL, 1μg/mL, 10μmol/L, and 2.5μmol/L individually after 48 h of culture. As2O3 with DXM, VP-16, MTX, BTZ, or SAHA showed remarkable antitu-mor efficacy compared with that of individual applications. Conclusion:As2O3 alone or combined with DXM, VP-16, MTX, BTZ, or SAHA significantly inhibited Hut-78 and Hut-102 cell growth in vitro. This study demonstrated that As2O3 with DXM, VP-16, MTX, BTZ, or SAHA presents synergistic antitumor effects on CTCL cells and may be an optimal regimen in clinical trials of CTCL.

Study on the mechanism of Arsenic Trioxide inducing the apoptosis of myelodysplastic syndrome cell line SKM-1 / 重庆医学

Objective To investigate the mechanism of AS2 O3 inducing the apoptosis of myelodysplastic syndrome(MDS) cell line SKM-1 .Methods SKM-1 cells were incubated with AS2 O3 ,and then the cellular morphology was observed ,flow cytometry was used to determine the apoptosis ,RT-PCR was used to detect the expressions of Bcl-2 ,Bax and caspase-3 mRNA .Results 0 .25、0 .50 μmol/L AS2 O3 could not markedly induce the apoptosis of SKM-1 cells (P>0 .05) .But 2 .00、8 .00、32 .00 μmol/L of AS2 O3 could obviously promote the apoptosis of SKM-1 cells .With the increase of the acting time and concentration of AS2 O3 ,the apoptosis rate increased ,too(P<0 .01) ,the expressions of anti-apoptotic gene Bcl-2 mRNA decreased (P<0 .01) ,the expressions of promoting apoptosis gene Bax and caspase-3 mRNA increased (P<0 .01 ,P<0 .05) .Conclusion 2 .00、8 .00、32 .00 μmol/L of AS2O3 may promote the apoptosis of SKM-1 cells through down-regulating the expression of Bcl-2 gene and up-regulating the ex-pressions of Bax and caspase-3 genes .

Induction of apoptosis in NB4 cells by decitabine alone and combination with As2O3 / 白血病·淋巴瘤

Objective To investigate the effect of methylation inhibitor decitabine (DAC) alone and combination with As2O3 on apoptosis of NB4 cells. Methods NB4 cells were treated with DAC, As2O3 and the combination of them in different concentrations. The cell proliferation was analyzed by MTT assay and the apoptosis of NB4 cells was detected by flow cytometry. Results Both DAC and As2O3 induced time and concentration-dependent cell death, in which the inhabitation rate were 12.18 %, 22.72 %, 35.54 %, respectively, after 24 h, 48 h, 72 h on treatment by DAC at 1 μmol/L and the inhibition rates were increased to 22.14 %, 31.18 %, 45.21 % by DAC at 1 μmol/L. The inhibition rates were 21.09 %, 32.43 %, 44.93 %, respectively, by treating with As2O3 0.5 μmol/L after 24 h, 48 h, 72 h, which were increased to 31.69 %, 41.12 % and 54.27 %, respectively after 24 h, 48 h, 72 h. The inhibition rates were significantly increased by using both DAC and As2O3 with significant differences (P <0.05). DAC and As2O3 in combination produced a greater inhibition of growth against NB4 cells (by treating with DAC 1 μmol/L + As2O3 0.5 μmol/L after 24 h, 48 h, 72 h, the inhibition rates were 42.10 %, 48.75 %, 60.78 %) (P <0.05). In each concentration group and control group the differences were statistically significant (P <0.05). The incubation for 48 h with As2O3 1 μmol/L alone or combined with DAC 2 μmol/L showed apoptosis cells by 5.8 % and 17.3 %. Conclusion Decitabine can significantly inhibit the proliferation of NB4 cells and the apoptosis with synergistic effectiveness can be found when Decitabine combination with As2O3.

Role of solasodine hydrochloride in AS2O3 induced HeLa cells apoptosis as well as its effect on cell telomerase activity in vitro / 中国地方病学杂志

Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.

Study on DNA damage of retinal pigment epithelium cell induced by arsenic trioxide in rabbit / 眼科研究

Background Researches have reported that arsenic trioxide (As_2O_3) is an effective method of treating solid tumor,and its mechanism is to inhibit the cellular proliferation and induce cellular apoptosis.However,the research on DNA damage caused by As_2O_3 is not clear.Objective This study is to investigate the roles of arsenic trioxide (As_2O_3) on DNA damage of retinal pigment epithelial cell in rabbit.Methods Retinal pigment epithelial cells were isolated from pigmented rabbits and cultured in vitro according to the method of Singh NP[2].Cultured cells were identified by cytokeratin.The third or fourth generation of cells were used to this study.50.00,5.00,2.00,1.00,0.50 and 0.25 μmol/L of As_2O_3 were added into medium respectively for 1 hour,and the same volume of PBS was added as negative control group.Cultured cells were exposed to ultraviolet (UV) rays with wave length of 254 nm for 10 minutes as positive control group.The cell vitality was detected by trypan-blue staining.The comet assay was applied to evaluate the DNA damage.SPSS 13.0 software was used for statistical analysis.Results The black granule were seen in cultured RPE cells.Cultured cells showed the positive brown-yellow particles in cytoplasm for cytokeratin with the positive rate over 90%.The morphology of the cells was similar among experimental group,UV irradiation group and PBS group under the inverted microscope.The normal cell activity was exhibited by trypan-blue staining in these three groups.Compared with PBS group,As_2O_3 caused obvious abnormality of DNA of RPE cell in a dose-dependent manner.Both the percentage of tail DNA and tail moment were gradually increased with the enhance of As_2O_3 concentration (F=88.548,P=0.000; F= 129.704,P=0.000).Significant differences also were found between different concentrations of As_2O_3 groups and UV irradiation group compared with PBS group (P＜0.05).As_2O_3 caused the obvious breaking up of DNA strands of RPE cells at the concentration of 50.00 μmol/L,but there was no statistical difference in spontaneous DNA damage of RPE cells between 0.50 and 0.25 μmol/L of As_2O_3 (P＞0.05).Conclusion As_2O_3 has a potential genotoxicity for rabbit retinal pigment epithelial cells in vitro.

Arsenic trioxide inhibits KM3 cell growth and induces apoptosis and potentiated by bortezomib / 中国病理生理杂志

AIM: To observe if there is a synergistical effect on induction of apoptosis when arsenic trioxide alone or combination with bortezomib in KM3 cells. METHODS: KM3 cells were treated with arsenic trioxide alone or combined with bortezomib, the numbers of viable cells were determined by trypan blue exclusion. Cell growth inhibition was examined by MTT method. The cells were simultaneously stained with annexin V-FITC and PI and apoptosis was determined by bivariate flow cytometry using a FACScan. Reverse trascriptional-PCR (RT-PCR) method was used to examine the change of p65 mRNA and Western blotting to measure the expression of protein p65, p-p65, caspase-3, -8, -9, and poly ADP-ribose polymerase (PARP). RESULTS: Arsenic trioxide inhibited the cell growth and induced apoptosis. The mechanism was responsible for the activation of caspase-mediated induction of apoptosis. A synergistic effect of combination with bortezomib on apoptosis was observed. CONCLUSION: Arsenic trioxide inhibits KM3 cell growth and induces apoptosis with a synergistical effect when cotreated with bortezomib.

Inhibitory effect of methylmercury on proliferation of human small cell lung cancer cells / 中国免疫学杂志

Objective:To investigate the inhibition of methylmercury on proliferation of human small cell lung cancer NCI-H446 and to compare with As_2O_3 and cisp latin.Methods:MTT chromatometry was adopted.Cultured NCI-H446 cells were incubated with different doses of methylmercury,As_2O_3,or cisplatin at 2.5,5,10,20,40 μmol/L.The optical density OD_(570) value were detected to calculate the inhibitory rate of proliferation.The nude mice model bearing tumor was constructed by subcutaneous injection with transplanting NCI-H446 cells.MMC was used at 10 mg/kg in administration with normal saline as the control.Tumor size was measured and tumor volume was calculated.Then the morphologic change was observed by HE stainning.Results:OD_(570) value of methylmercury groups were lower than that of the control groups at any doses and the difference was significant from 2.5 μmol/L group(P<0.05).The difference of the OD_(570) value of methylmercury groups under the same concentration among the different times were significant (P＜0.05);OD_(570) value of As_2O_3 and cisplatin groups were also lower than that of the control groups.There was no significant difference with MMC and cisplatin.OD_(570) value of MMC and DDP groups were higher than that of As_2O_3 groups.The volume of the tumor of the experiment groups was smaller than that of normal saline group,and the necrocytosis was more than that of normal saline group by HE stainning.Conclusion:Methylmercury could inhibit the proliferation of human small cell lung cancer NCI-H446,and the effect is both time- and dose-dependence.At the same doses,this effect may be equal to cisplatin's and stronger than As_2O_3's.

Effect of arsenic trioxide on growth of a heterologous graft model for human breast cancer in nude mice / 国际外科学杂志

Objective To study the inhibitory effect of arsenic trioxide(AS2O3)on the tumor growth of breast cancer line MDA-MB-435s implanted subcutaneously in nude mice and its mechanism.Methods BALB/C-nu/nu nude mice were subcutaneously injected with MDA-MB-435s breast cancer cells that are ER-positive,and treated with intraperitoneal injection of AS2O3 and DDP in different concentrations.The implanted tumor was weighed,and tumor inhibition rates were calculated.The expression of PTEN and Caspase-7 induced by AS2 03 were examined by immunohistochemical method.Results The growth of implanted tumor was markedly inhibited with DDP,low dose and high dose AS2O3,and the inhibitory rates were 48.68%,32.80%,66.67%respectively.The immunohisto chemical staining showed that the number of PTEN and Caspase-7 protein increased markedly(P＜0.05).Conclu sions AS2O3 inhibits the growth of human breast cancer cell implanted tumor.The molecular mechanism of AS2O3 on induction of apoptosis of breast cancer cells may be throush increasing the expression of PrrEN and Caspase-7(P ＜0.05).

Vascular shutdown effects by tetraarsenic oxide in TC-1 cells implanted C57BL/6 mice / 대한산부인과학회지

OBJECTIVE: Arsenic trioxide (As2O3) is known to have potent anti-vascular activity and significantly suppress solid tumor growth. The present study was conducted to investigate the vascular shutdown effects of a novel arsenic compound, tetraasrsenic oxide (As4O6), in comparison with As2O3 using cervical cancer animal model. METHODS: Mice tumor challenge model was used C57BL/6 mice transplanted with TC-1 cells. After the growth of tumors was reached up 200~250 mm3, mice were divided into 3 groups randomly for control and treatment of either As2O3 or As4O6. As2O3 and As4O6 was treated by i.p. injection. The tumor size was caliperated in twice for weeks and anti-vascular effect were assessed by Evans blue extraction assay and Hoechst 33342 staining. In tumor tissue, histopathological feaure was obserevd by hematoxylin and eosin (H&E) staining. RESULTS: In mice treated with either As2O3 and As4O6 (i.p.), both of As2O3 and As4O6 was significantly suppressed the tumor growth compared with control group. Moreover, effect of As4O6 is more pronounced. These tumor growth inhibition is led to the massive necrosis and vacular shutdown in tumor tissue. CONCLUSION: This study suggests that As4O6 may have potential anticancer activity via vascular shutdown in C57BL/6 mice transplanted with TC-1 cells.

Effect of telomerase-antisense DNA on apoptosis of hepatoma cells induced by arsenic trioxide / 中国肿瘤生物治疗杂志

Objective: To observe the effect of telomerase-antisense DNA on apoptosis of hepatoma cells induced by arsenic trioxide, in an effort to look for a new anti-hepatic cancer agent with high efficiency, low cytotoxicity. Methods: We designed and synthesized a 20nt telomerase-antisense DNA targeting telomerase template and observed its influence on the telomerase activity of hepatoma cells. H-E staining, flow cytometry, and DNA agarose electrophoresis were used to study the preventive effect of telomerase-antisense DNA on hepatoma cells apoptosis induced by arsenic trioxide. Flow cytometry was used to detect the expression of Fas, Fas-L, and bcl-2. Results: Telomerase-antisense DNA (5 ?mol/L) effectively inhibited the telomerase activity of hepatoma cells after 24 hours (P

Effect of As_2O_3 on mouse bone marrow hemopoietic microenvironment / 吉林大学学报(医学版)

Objective To investigate the effect of As_2O_3 on normal mouse bone marrow stromal cells(BMSCs).Methods BMSCs and colony-forming unit-granulocyte macrophage (CFU-GM) were cultured in vitro to observe the quantitative changes of colony-forming unit-fibroblast(CFU-F) and CFU-GM after BMSCs were treated with different concentrations of As_2O_3 respectively;the quantitative changes of CFU-GM supported by BMSCs and granulocyte colony stimulating factor(G-CSF) and IL-11 secreted by BMSCs were also observed after the progenitor cells supported by BMSCs were treated with As_2O_3 for 3,5 and 7 d,respectively.Results The productions of CFU-F and CFU-GM supported by BMSCs were decreased obviously when treated with 7 ?mol?L~(-1) As_2O_3,also the levels of G-CSF and IL-11 secreted by BMSC(P

Preliminary Study on Apoptosis of Human Gastric Carcinoma Cell line MKN45 Induced by Arsenic Trioxide and its Molecular Mechanism / 医学研究杂志

Objective To investigate the possibility of human gastric carcinoma cells apoptosis induced by arsenic trioxide and its mechanism.Methods After treatment with arsenic trioxide, the cytotoxicity to human gastric carcinoma cells MKN45 was quantified using trypan blue exclusion, and IC 50 was determined. Apoptotic cells were detected with flow cytometry, DNA cytofluorometry, DNA electrophoresis. Results Arsenic trioxide inhibited the growth of human gastric carcinoma cells MKN45 in a dose-dependent manner in a certain range of dose with a IC 50 of (11.05?0.25)?mol/L; Apoptotic peak, characteristic morphologic features of apoptosis and DNA ladder were observed in human gastric carcinoma cells MKN45 treated with 1~10 ?mol/L arsenic trioxide. Conclusions Arsenic trioxide can induce apoptosis of human gastric carcinoma cells MKN45, suggesting a great potential in the treatment of gastric carcinoma.