Study on the Antitumor Effects of Low-energy Laser Irradiation Combined with Cyclophosphamide

Systematic experiments about the antitumor effects of low energy laser irradiation combined with the traditional antitumor medicine of cyclophosphamide were conducted using the experimental model of mouse S180 ascites sarcoma.The three groups of tumor bearing mice were irradiated upon the inner corners with the dosages of 11 00,14 67 and 22 00 J·cm -2 LELI respectively,and injected with CYT intraperitoneally to observe the changes of the survival time,the ascites growth speed,and the kinetic changes of immune functions.The survival times of the three groups of CYT/LELI combination were obviously longer than those of the tumor and CYT control groups.Correspondingly,the amounts of ascites,tumor cells densities and total tumor cells in CYT/LELI groups decreased significantly,while the death ratio of the tumor cells increased.Comparatively,the group of 22 00 J·cm -2 LELI combined with CYT showed the most ideal antitumor effects,and the life prolongation ratio was up to 53 20%.

ANTITUMOR EFFECTS OF WICK OF SUN-FLOWER STEM IN MICE

A well-known Chinese traditional drug, decoction of wick of sun-flower stem, has been used to treat cancer and certain other diseases.1,2 Animal experiments in mice showed that transplantation of Sarcoma 180 (S180) and of Uterus cervical cancer 14 (U14) were signlflcanly inhibited by this decoction. The Inhibition rates of tumor weight were 33 -81 % and in about 6 - 20% of treated mice, complete tumor regression has been demonstrated. So tar no acute toxic side-effect was noted. This is in contrast to most of the antitumor chemltherapeutic drugs known to produce different degrees of toxlcity or induce Immunosuppression as a side effect. For this reason, we have undertaken the present study.

Purification,structural elucidation,and in vitro antitumor effects of novel polysaccharides from Bangia fuscopurpurea

A water-soluble polysaccharide,designated BFP-3,was isolated from Bangia fuscopurpurea by hot water extraction,anion-exchange,and size-exclusion chromatography and tested to determine its antitumor activity.The structural characteristics of BFP-3 were investigated by chemical and spectroscopic methods,including partial acid hydrolysis,methylation analysis,one-and two-dimensional nuclear magnetic resonance,and gas chromatography-mass spectrometry.The results showed that BFP-3 was mainly comprised of rhamnose,arabinose,mannose,glucose,and galactose.Moreover,the weight-average molecular weight of BFP-3 was estimated to be approximately 333 kDa.The backbone of BFP-3 was primarily composed of repeating 5-α-l-Araf-1→(4-α-d-Glcp-1)_(4)→4,6-β-d-Manp-1 units,and the side chains consisted of repeatingβ-d-Galp-1→(4-β-d-Galp-1)_(4)→4,6-β-d-Galp-1→3,4-α-l-Rhap,β-l-Arap-1→(3-β-d-Galp-1)_(3),andβ-l-Arap-1 units.Counting Kit-8 assays revealed that BFP-3 significantly inhibited the proliferation of A2780,COC1,SKOV3,HO-8910,and OVCAR3 ovarian cancer cells in vitro,indicating that BFP-3 could have potential applications in the treatment of ovarian cancer.

Antitumor Effects of Ethanol Extract from Ventilago leiocarpa Benth on Sarcoma 180 Tumor-Bearing Mice and Possible Immune Mechanism

Objective To explore the antitumor effects of ethanol extract from Ventilago leiocarpa Benth(EEVLB)on sarcoma 180(S180)tumor-bearing mice and the potential mechanism.Methods Sixty mice were randomly assigned to 6 groups according to a random number table:normal group,model group,5-fluorouracil(5-FU)group(0.02 g·kg^(−1)),and high-,medium-,low-dose EEVLB groups(100,84,and 56 g of raw material·kg^(−1)body weight,respectively),with 10 mice each group.All treatments were given once daily for 10 consecutive days.Effects of EEVLB on inhibiting tumor growth and immune function in mice were evaluated among all groups after the treatments by detecting tumor inhibition rate,organ index,serum levels of interleukin(IL)-2,-6,-10,CD3^(+)CD4^(+)T lymphocytes,CD4^(+)/CD8^(+)ratio,caspase-3 and Bcl-2.Results EEVLB with different concentrations achieved inhibition of tumor growth in vivo,wherein the high-dose group showed the most significant reduction in tumor weight and increased apoptosis of tumor cells(P<0.05).In addition,both net weight gain and spleen index of mice showed uptrend in EEVLB treatment groups(P<0.05).Besides,serum levels of IL-2 and IL-6,percentages of CD3^(+)CD4^(+)T lymphocytes and ratio of CD4^(+)/CD8^(+)in peripheral blood were elevated in high-and medium-dose EEVLB groups compared with the model group(P<0.05).Also,upregulation of caspase-3 and downregulation of Bcl-2 were observed at protein levels in the high-dose EEVLB group(P<0.01).Conclusions EEVLB exhibits promising antitumor activity in vivo.This effect might be due to activation of apoptotic signaling pathway,increase of cytokine levels and enhancement of immune function in tumor-bearing mice.

Preparation, characterization, pharmacokinetics and anticancer effects of PEGylated β-elemene liposomes

Objective:This study aimed to develop a new polyethylene glycol(PEG)ylatedβ-elemene liposome(PEG-Lipo-β-E)and evaluate its characterization,pharmacokinetics,antitumor effects and safety in vitro and in vivo.Methods:The liposomes were prepared by ethanol injection and high-pressure micro-jet homogenization.Characterization of the liposomes was conducted,and drug content,entrapment efficiency(EE),in vitro release and stability were studied by ultra-fast liquid chromatography(UFLC)and a liquid surface method.Blood was drawn from rats to establish the pharmacokinetic parameters.The anticancer effect was evaluated in a KU-19-19 bladder cancer xenograft model.Histological analyses were performed to evaluate safety.Results:The PEG-Lipo-β-E showed good stability and was characterized as 83.31±0.181 nm in size,0.279±0.004 in polydispersity index(PDI),-21.4±1.06 mV in zeta potential,6.65±0.02 in pH,5.024±0.107 mg/mL inβ-elemene(β-E)content,and 95.53±1.712%in average EE.The Fourier transform infrared spectroscopy(FTIR)and differential scanning calorimetry(DSC)indicated the formation of PEG-Lipo-β-E.Compared to elemene injection,PEG-Lipo-β-E demonstrated a 1.75-fold decrease in clearance,a 1.62-fold increase in half-life,and a 1.76-fold increase in area under the concentration-time curves(AUCs)from 0 hour to 1.5 hours(P<0.05).PEG-Lipo-β-E also showed an enhanced anticancer effect in vivo.Histological analyses showed that there was no evidence of toxicity to the heart,kidney,liver,lung or spleen.Conclusions:The present study demonstrates PEG-Lipo-β-E as a new formulation with ease of preparation,high EE,good stability,improved bioavailability and antitumor effects.

Excellent effects and possible mechanisms of action of a new antibody–drug conjugate against EGFR-positive triple-negative breast cancer

Background:Triple-negative breast cancer(TNBC)is the most aggressive subtype and occurs in approximately 15%–20%of diagnosed breast cancers.TNBC is characterized by its highly metastatic and recurrent features,as well as a lack of specific targets and targeted therapeutics.Epidermal growth factor receptor(EGFR)is highly expressed in a variety of tumors,especially in TNBC.LR004-VC-MMAE is a new EGFR-targeting antibody–drug conjugate produced by our laboratory.This study aimed to evaluate its antitumor activities against EGFR-positive TNBC and further studied its possible mechanism of antitumor action.Methods:LR004-VC-MMAE was prepared by coupling a cytotoxic payload(MMAE)to an anti-EGFR antibody(LR004)via a linker,and the drug-to-antibody ratio(DAR)was analyzed by HIC-HPLC.The gene expression of EGFR in a series of breast cancer cell lines was assessed using a publicly available microarray dataset(GSE41313)and Western blotting.MDA-MB-468 and MDA-MB-231 cells were treated with LR004-VC-MMAE(0,0.0066,0.066,0.66,6.6 nmol/L),and the inhibitory effects of LR004-VC-MMAE on cell proliferation were examined by CCK-8 and colony formation.The migration and invasion capacity of MDA-MB-468 and MDA-MB-231 cells were tested at different LR004-VCMMAE concentrations(2.5 and 5 nmol/L)with wound healing and Transwell invasion assays.Flow cytometric analysis and tumorsphere-forming assays were used to detect the killing effects of LR004-VC-MMAE on cancer stem cells(MDA-MB-468 and MDA-MB-231 cells).The mouse xenograft models were also used to evaluate the antitumor efficacy of LR004-VC-MMAE in vivo.Briefly,BALB/c nude mice were subcutaneously inoculated with MDA-MB-468 or MDAMB-231 cells.Then they were randomly divided into 4 groups(n=6 per group)and treated with PBS,naked LR004(10 mg/kg),LR004-VC-MMAE(10 mg/kg),or doxorubicin,respectively.Tumor sizes and the body weights of mice were measured every 4 d.The effects of LR004-VC-MMAE on apoptosis and cell cycle distribution were analyzed by flow cytometry.Western blotting was used to detect the effects of LR004-VC-MMAE on EGFR,ERK,MEK phosphorylation and tumor stemness marker gene expression.Results:LR004-VC-MMAE with a DAR of 4.02 were obtained.The expression of EGFR was found to be significantly higher in TNBC cells compared with non-TNBC cells(P<0.01).LR004-VC-MMAE inhibited the proliferation of EGFRpositive TNBC cells,and the ICvalues of MDA-MB-468 and MDA-MB-231 cells treated with LR004-VC-MMAE for 72 h were(0.13±0.02)nmol/L and(0.66±0.06)nmol/L,respectively,which were significantly lower than that of cells treated with MMAE[(3.20±0.60)nmol/L,P<0.01,and(6.60±0.50)nmol/L,P<0.001].LR004-VC-MMAE effectively inhibited migration and invasion of MDA-MB-468 and MDA-MB-231 cells.Moreover,LR004-VC-MMAE also killed tumor stem cells in EGFR-positive TNBC cells and impaired their tumorsphere-forming ability.In TNBC xenograft models,LR004-VC-MMAE at 10 mg/kg significantly suppressed tumor growth and achieved complete tumor regression on day 36.Surprisingly,tumor recurrence was not observed until the end of the experiment on day 52.In a mechanistic study,we found that LR004-VC-MMAE significantly induced cell apoptosis and cell cycle arrest at G/M phase in MDAMB-468[(34±5)%vs.(12±2)%,P<0.001]and MDA-MB-231[(27±4)%vs.(18±3)%,P<0.01]cells.LR004-VC-MMAE also inhibited the activation of EGFR signaling and the expression of cancer stemness marker genes such as Oct4,Sox2,KLF4 and EpCAM.Conclusions:LR004-VC-MMAE showed effective antitumor activity by inhibiting the activation of EGFR signaling and the expression of cancer stemness marker genes.It might be a promising therapeutic candidate and provides a potential therapeutic avenue for the treatment of EGFR-positive TNBC.

PLASMID VACCINE ENCODING HN GENE FROM NEWCASTLE DISEASE VIRUS HAS MARKED ANTITUMORAL EFFECT IN VITRO

Objective: To explore the antitumor effects of hemaagglutinin-neuraminase gene (HN gene) from Newcastle disease virus. Methods: Plasmid vaccine of pIRHN was constructed and transfected into HeLa cells. The expression of HN was analyzed by Western blot analysis, and the mode of cell death was detected by fluorescence microscope, gel electrophoresis and TUNEL assay and the expression of p53 and bcl-2 was also analyzed in transfected Hela cells. The effect of pIRHN on sialic acid contents in the Hela cell was examined. Results: pIRHN nucleic acid vaccines could be expressed in eukaryotic cell. pIRHN could induce apoptosis after HeLa cells were transfected. The effect of antitumor responses of pIRHN was correlated with the contents of sialic acid in tumor cells, and there was no prominent evidence for the relatedness of the antitumor effect with the expression of p53 and bcl-2. Conclusion: pIRHN may become a new antitumor biological agent.

ANTITUMOR EFFECT OF ACTINIDIA CHINENSIS POLYSACCHARIDE ON MURINE TUMOR

A new polysaccharide compound (ACPS-R) has recently been isolated from the root of Actinidia Chinensis Planch. When given inttaperitoneally to the transplantable tumor bearing mice at dose of 75-125 mg/kg, the tumor inhibition rate was more than 88.8% in Ehrilich ascitic cancer (EAC) or ascitic from of hepatoma (HepA) and more than 49.6% in solid hepatoma (HepS). The treatment effect of ACPS-R on EAC at dose of 80-100 mg kg and 125 mg/kg were comparable to that of cyclophosphamide at dose of 15 mg/kg and 22.5 mg/kg, respectively. ACPS-R could also prolong the life of EAC- or P388-bearing mice, and increase the percentage of EAC-free mice. In addition, when ACPS-R was used in combination with 5-Fu, the antitumor effect was enhanced as compared with 5-Fu alone. A marked increase in cAMP levels end cAMP cGMP ratio of spleen of EAC-bearing mice were observed after treatment of ACPS-R. The increase of both para meters nearly reached the normal levels of healthy mice. The increases of cAMP, cAMP cGMP and tumor remission had statistical significance. It showed an intermediate inhibitory effect of ACPS-R on DNA synthesis by incorporating 3H-TdR into EAC cells.The results indicated that ACPS-R acts as a new antitumor polysaccharide, and the treatment effect of Actinidia root in folk medicine is probably related to ACPS-R.

EFFECT OF ANTITUMOR DRUGS ON THE ACTIVITY OF DNA TOPOISOMERASE I

The activity of DNA topoisomerase Ⅱ prepared from either normal or tumor tissues were compared. It was found that the unknotting activity of the enzyme in malignant tumor cells was higher than that in normal cells. We selected some antitumor drugs including Chinese traditional medicine, and observed their effects on the unknotting activity of topoisomerase Ⅱ. The results showed that inhibition of the unknotting activity of the enzyme required very low concentrations of drugs, but much higher concentrations were required for other tested. Some antitumor drugs had no effect on the enzyme were also proved. It is interesting that carrageenan, an antiviral drug, strongly blocked the unknotting activity although its antitumor activity has not been reported.

THE ANTITUMOR ACTIVITY OF LYMPHOTOXIN AND THE EFFECT OF INTERLEUKIN-2 ON ITS FUNCTION

The effect of lymphotoxin (LT)-containing supernatant produced by lectin-stimulated human lymphocytes on tumor cells and the relation between interleukin-2 (IL-2) and LT were studied in this article. Results showed that LT-containing superna-tants had cytotoxicities on many different kinds of tumor cells from human and mice, that actinomycin D increased the LT activities on target cells and that IL-2 had the ability to increase the cytotoxicity of human PBMC on tumor cells, after being treated with LT, the target cells were more easy to kill by PBMC as well.

Pharmacological Action and Molecular Mechanism of Amentoflavone

Amentoflavone(AMF)is a natural active substance extracted from Chinese herbal medicine Selaginella tamariscina,and has good anti-inflammatory,antitumor,hypoglycemic and neuroprotective pharmacological effects.This paper reviews the pharmacological action and mechanism of AMF,in order to lay a theoretical foundation for in-depth research and drug development of AMF.

THE ANTICANCER EFFECT AND ANTI－DNA TOPOISOMERASE II EFFECT OF EXTRACTS OF CAMELLIA PTILOPHYLLA CHANG AND CAMELLIA SINENSIS

The cytotoxic effect of extract of camellia ptilophyllachang(ECPC) and extract of camellia sinensis(ECS) onHeLa cell line, poorly differentiated nasopharyngealcarcinoma cell line(CNE2) and gastric cancer cell line(MGC-803 ) in vitro was studied using MIT assay method.The results showed that ECPC and ECS possessed significantcytotoxic effect on above three cell lines. The anticancer testin mice showed that ECPC had marked inhibitory effectagainst Ehrlich solid carcinoma(ESC) with inhibition ratesof 17. 8 48. 3% and with inhibition rates of 28. 3-54. 5% against reticular cell sarcoma(L2), and that ECShad inhibition rates of 31 . 5 -49. 4 % against ESC and 35. 8- 50% against L2. These two extracts had only marginalinhibitory effect against sarcoma- 180. The unknottingactivity of DNA topoisomerase II was inhibited completelyby ECPC and ECS at the concentration of 50 μg/ mlsuggesting that DNA topoisomerase II might be a targetenzyme of these two extracts.

Progress on the Study of the Anti-Tumor Effect of Emodin

Cancer is a disease caused by the loss of normal cell regulation and excessive proliferation. At present, there are a lot of anticancer drugs, but most of them are not ideal, with sereve side effects. Besides, during the treatment, the patients feel very bad, and they are always in great pain. Emodin is a natural anthraquinone derivative found in a variety of herbal preparations. Many studies have shown that emodin has a significant therapeutic effect on lung cancer, liver cancer, breast cancer, and so on. After reviewing a large amount of literatures, this paper summarizes the research progresses of emodin anticancer in the past five years, in order to provide theoretical basis for further development and utilization of emodin and its metabolites.

In vitro antitumor effect of cucurbitacin E on human lung cancer cell line and its molecular mechanism

Cucurbitacin E(CuE) is previously reported to exhibit antitumor effect by several means.In this study, CuE acted as a tyrosine kinase inhibitor interfering with the epidermal growth factor receptor/mitogen-activated protein kinase(EGFR/MAPK) signaling pathway and subsequently induced apoptosis and cell cycle arrest in non-small-cell lung cancer(NSCLC) cell line A549.The apoptosis regulators, cleaved Caspases-3 and Caspases-9, were observed to be increased with the treatment of CuE.The activated transcription factor STAT3 and the apoptosis inhibitor protein survivin were also observed to be reduced.The cell cycle regulators, CyclinA2, cylinB1, CyclinD1 and CyclinE, were also investigated and the results suggested that the cell cycle was arrested at G1/G0 phase.Treatment of CuE also altered the existence status of most of the participants in the EGFR/MAPK signaling.Phosphorylation of EGFR enhanced significantly, leading to the alteration of members downstream, either total amount or phosphorylation level, notably,MEK1/2 and ERK1/2.Moreover, the results of molecular simulation brought an insight on the interaction mechanism between CuE and EGFR.In summary, CuE exhibited anti-proliferative effect against A549 cells by targeting the EGFR/MAPK signaling pathway.

Enhancement of Antitumor Effect of Tegafur/Uracil(UFT) plus Leucovorin by Combined Treatment with Protein-Bound Polysaccharide,PSK,in Mouse Models

We evaluated the antitumor effect of combined therapy with tegafur/uracil （UFT） plus leucovorin （LV） （UFT/LV） and protein-bound polysaccharide, PSK, in three mouse models of transplantable tumors. UFT/LV showed antitumor effect against Meth A sarcoma, and the antitumor effect was enhanced when PSK given concomitantly. UFT/LV showed antitumor effect to Lewis lung carcinoma and PSK alone also showed antitumor effect at high dose, but a combination of UFT/LV and PSK resulted in no enhanced antitumor effect. Colon 26 carcinoma was weakly responsive to UFT/LV, and no enhancement of antitumor effect was found even PSK was used in combination. In conclusion, while the effect of PSK varies depending on tumor, combined use of UFT/LV and PSK may be expected to augment the antitumor effect.

New possible silver lining for pancreatic cancer therapy:Hydrogen sulfide and its donors

As one of the most lethal diseases,pancreatic cancer shows a dismal overall prognosis and high resistance to most treatment modalities.Furthermore,pancreatic cancer escapes early detection during the curable period because early symptoms rarely emerge and specific markers for this disease have not been found.Although combinations of new drugs,multimodal therapies,and adjuvants prolong survival,most patients still relapse after surgery and eventually die.Consequently,the search for more effective treatments for pancreatic cancer is highly relevant and justified.As a newly re-discovered mediator of gasotransmission,hydrogen sulfide(H2S)undertakes essential functions,encompassing various signaling complexes that occupy key processes in human biology.Accumulating evidence indicates that H2S exhibits bimodal modulation of cancer development.Thus,endogenous or low levels of exogenous H2S are thought to promote cancer,whereas high doses of exogenous H2S suppress tumor proliferation.Similarly,inhibition of endogenous H2S production also suppresses tumor proliferation.Accordingly,H2S biosynthesis inhibitors and H2S supplementation(H2S donors)are two distinct strategies for the treatment of cancer.Unfortunately,modulation of endogenous H2S on pancreatic cancer has not been studied so far.However,H2S donors and their derivatives have been extensively studied as potential therapeutic agents for pancreatic cancer therapy by inhibiting cell proliferation,inducing apoptosis,arresting cell cycle,and suppressing invasion and migration through exploiting multiple signaling pathways.As far as we know,there is no review of the effects of H2S donors on pancreatic cancer.Based on these concerns,the therapeutic effects of some H2S donors and NO-H2S dual donors on pancreatic cancer were summarized in this paper.Exogenous H2S donors may be promising compounds for pancreatic cancer treatment.

New understanding of the low-dose radiation-induced hormesis

Dr.Liu Shuzheng as a well-known educator and radiobiologist has started to work on the physiological benefits of low-dose radiation since 1970s.His research on the distinct effects of low-dose radiation from those of high-dose radiation has nationally and internationally impacted radiobiology,medicine and oncology.Therefore,here we briefly review the research from his research team from early years to last two decades.Two important facts related to LDR biological effects are the hormesis and adaptive response,both have been the main focuses of Dr.Liu Shuzheng early research.In the last couple of decades Dr.Liu Shuzheng have further investigated how to apply these important mechanisms into clinical translational research,such as the potential to application of LDRhormesis to enhance tumor’s chemo-or radio-therapy and LDR-induced adaptive response to protect normal tissue from chemo-or radio-therapy related side toxic effects.Therefore,Dr.Liu has provided important guidance and role model for his trainees in the continue of their research career in radiation biology,medicine and oncology.

Inhibition of Yuyihe Powder on Tumor Growth in Mice Models of Sarcoma 180

Objective To explore the antitumor effect of Yuyihe Powder（Yu Yi He San,YYHS） and its antitumor mechanism.Methods After treatment,tumor weight,immune apparatus weight,the life span of transplanted animals,spleen lymphocyte proliferation assays,and IL-2 concentration in mouse serum were recorded or detected.Results YYHS showed strong antitumor ability.Compared with control group,mid-dose YYHS（1.0g/kg） could inhibit the tumor growth,prolong the life span of S180-bearing mice to some extent,significantly increase the thymic and splenic indices of S180 mice,and strongly promote the secretion of IL-2 in blood;The inhibitory rate on tumor growth and life prolongation rate were 37.1%and 38.37%,respectively.Conclusion YYHS could not only significantly inhibit the growth of S180 cells,but also markedly prolong the survival time of S180 bearing mice.The mechanism of antitumor effect could obviously enhance immunologic function of the S180 bearing mice to inhibit the growth of S180 cells.

Cerium(Ⅳ)ammonium nitrate(CAN)-mediated regioselective synthesis and anticancer activity of 6-substituted 5,8-dimethoxy-1,4-naphthoquinone

6-Substituted 5,8-O-dimethyl-1,4-naphthoquinones（6-DMNQ）,the promising anticancer scaffolds,were selectively generated by oxidative demethylation of 2-substituted 1,4,5,8-tetramethoxynaphthalenes with CAN in EtOAc/H2O in comparatively high yields.An interesting finding was that apart from the reported electron-withdrawing effects of substituents on position 2 of naphthalene ring,regioselective synthesis of 6-DMNQ was largely dependent on the steric effects in CAN-mediated oxidation.The selective cytotoxicities of 6-DMNQ from the in vitro cell-based assays were exhibited between the cancer cells and normal cells.Moreover,most of sulfur-containing 6-DMNQ derivatives displayed better anticancer activities than the corresponding oxygen-containing ones,which could provide an available strategy for the design of 6-DMNQ derivatives as potential anticancer agents.