Immunogenic Cell Death Photothermally Mediated by Erythrocyte Membrane-Coated Magnetofluorescent Nanocarriers Improves Survival in Sarcoma Model.

Inducing immunogenic cell death (ICD) during cancer therapy is a major challenge that might significantly improve patient survival. The purpose of this study was to develop a theranostic nanocarrier, capable both of conveying a cytotoxic thermal dose when mediating photothermal therapy (PTT) after its intravenous delivery, and of consequently inducing ICD, improving survival. The nanocarrier consists of red blood cell membranes (RBCm) embedding the near-infrared dye IR-780 (IR) and camouflaging Mn-ferrite nanoparticles (RBCm-IR-Mn). The RBCm-IR-Mn nanocarriers were characterized by size, morphology, surface charge, magnetic, photophysical, and photothermal properties. Their photothermal conversion efficiency was found to be size- and concentration-dependent. Late apoptosis was observed as the cell death mechanism for PTT. Calreticulin and HMGB1 protein levels increased for in vitro PTT with temperature around 55 °C (ablative regime) but not for 44 °C (hyperthermia), suggesting ICD elicitation under ablation. RBCm-IR-Mn were then intravenously administered in sarcoma S180-bearing Swiss mice, and in vivo ablative PTT was performed five days later. Tumor volumes were monitored for the subsequent 120 days. RBCm-IR-Mn-mediated PTT promoted tumor regression in 11/12 animals, with an overall survival rate of 85% (11/13). Our results demonstrate that the RBCm-IR-Mn nanocarriers are great candidates for PTT-induced cancer immunotherapy.

Oxidative stress and histopathological changes in several organs of mice injected with biogenic silver nanoparticles.

The widespread use of silver nanoparticles (AgNPs) requires a study of their safety. The aim of the present study was to assess the levels of oxidative stress markers and histopathological changes in the experimental model of sarcoma S-180 of outbred mice caused by biogenic AgNPs. AgNPs were synthesized using 50% ethanol extract of Ocimum araratum leaves that was standardized for rosmarinic acid content. The effects of AgNPs were tested on chemiluminescence (ChL), malonic dialdehyde (MDA) content and activity of superoxide dismutase (SOD) in healthy and experimental model of sarcoma S-180 mice. It was shown that, under the influence of AgNPs, the intensity of ChL decreased, in contrast with control groups (with the exception of the hepatocytes of animals with transplanted sarcoma). The presence of AgNPs leads to the decrease of MDA in the tissues of healthy mice and to a slight increase of MDA content in the tumour and kidney tissues. AgNPs neutralize the activity of SOD in kidney tissue samples in animals with transplanted sarcoma, and in tumour tissue, they reduce SOD activity by three times. The results of the histological analysis indicate that AgNPs not only cause the destruction of tumour tissue but also lead to structural changes in hepatocytes and nephrons, which can affect the function of these organs. AgNPs are potential agents for antitumor therapy. Future studies are needed using biocompatible non-toxic NPs that meet the requirement for these drugs.

Transformation Mechanism of Rare Ginsenosides in American Ginseng by Different Processing Methods and Antitumour Effects.

The mechanism by which ginsenosides from Panax quinquefolium L. transform into rare saponins by different processing methods and their antitumour effects have yet to be fully elucidated. Our study aimed to detect the effect of amino acids and processing methods on the conversion of ginsenosides in American ginseng to rare ginsenosides, using 8 monomeric ginsenosides as substrates to discuss the reaction pathway and mechanism. S180 tumour-bearing mice were established to study the antitumour effects of American ginseng total saponins (AGS-Q) or American ginseng total saponins after transformation (AGS-H) synergistic CTX. The results showed that aspartic acid was the best catalyst, and the thermal extraction method had the best effect. Under the optimal conditions, including a reaction temperature of 110°C, an aspartic acid concentration of 5%, a reaction time of 2.5 h and a liquid-solid ratio of 30 mL/g, the highest conversion of Rk1 and Rg5 was 6.58 ± 0.11 mg/g and 3.74 ± 0.05 mg/g, respectively. In the reaction pathway, the diol group saponins participated in the transformation process, and the triol group saponins basically did not participate in the transformation process. AGS-Q or AGS-H synergistic CTX, or AGS-H synergistic CTX/2 could significantly increase the tumour inhibition rate, spleen index and white blood cell count, had a significant upregulation effect on IL-2 and IL-10 immune cytokines; significantly restored the ratio of CD4+/CD8+; and significantly inhibited the level of CD4+CD25+. AGS-Q or AGS-H synergistic with CTX or CTX/2 can significantly upregulate the expression of Bax and cleaved-Caspase-3 and inhibit the expression of antiapoptotic protein Bcl-2. AGS synergistic CTX in the treatment of S180 tumour-bearing mice can improve the efficacy and reduce toxicity.

Study on Extraction Process of Root of Henry Wood Betony Polysaccharides and Their Antitumor Activity against S180.

We optimized the hot water extraction of polysaccharides from the root of Henry wood betony (RHWPs) using a uniform test and explored their anti-tumor activities in vitro and in vivo. The optimal extraction conditions were as follows: 40 min extraction time, liquid/solid ratio 30 mL/g, 100 min soaking time, two extraction cycles, 100% ethanol concentration, and extraction temperature of 80 °C. The molecular weight distribution of RHWPs with MWs was 228,600 g/mol and 5001 g/mol. The IR spectrum further indicated that RHWPs are acidic polysaccharides containing pyranose and furan rings. The main monosaccharides found in RHWPs were mannose, ribose, l-rhamnose monohydrate, glucuronic acid, galacturonic acid, glucose, galactose, xylose, arabinose, and fucose. RHWPs inhibited the proliferation of S180 tumor cells and induced apoptosis in vitro. Oral administration of RHWPs to tumor-bearing mice significantly inhibited the growth of the S180 xenografts, accelerated apoptosis in tumor cells, and expanded the necrotic regions. Furthermore, RHWPs also markedly increased the levels of TNF-α, IFN-γ, and IL-2 in the sera of tumor-bearing mice, and activated immune cells such as lymphocytes, NK cells, and macrophages, thereby inducing tumor cell apoptosis. Taken together, RHWPs are a promising anti-tumor agent that ought to be explored further.

Anti-tumor activity and immunogenicity of a succinoglycan riclin.

Natural polysaccharides have attracted considerable interests due to diverse biological activities. Succinoglycan is an extracellular polysaccharide produced by most Agrobacterium strains. Here, we confirmed riclin was a typical succinoglycan by NMR and methylation analysis, and investigated the antitumor effects of riclin in sarcoma 180 tumor-bearing mice. The results showed that riclin inhibited the tumor growth significantly as well as cyclophosphamide (CTX). While CTX caused serious damage to spleen structure, riclin increased the spleen index and promoted lymphocytes proliferation in peripheral blood, spleen and lymph nodes. Riclin decreased splenocytes apoptosis as evidenced by alterations of B-cell lymphoma-2 family proteins and Cleaved Caspase-3 protein. Moreover, 1H nuclear magnetic resonance (NMR)-based metabolomics analysis revealed that riclin partially altered the metabolic profiles of splenocytes. In conclusion, riclin is a succinoglycan that performed strong immunogenicity and suppressed sarcoma growth in mice. Succinoglycan riclin could be a potential antitumor agent for functional food and pharmaceutical purpose.

Intermittent hypoxia induces tumor immune escape in murine S180 solid tumors via the upregulation of TGF-ß1 in mice.

PURPOSE: Studies have shown that intermittent hypoxia (IH) alters host immune functions and promotes tumor growth. However, the relevant mechanisms of these effects have not been completely elucidated. We hypothesized that IH promotes the growth of tumors by changing cytokine levels in the tumor microenvironment and inducing immune escape. METHODS: Sarcoma-180 (S180) solid tumor cells were injected into the right flank of Kunming mice. The mice were then randomly divided into the IH and room air (RA) groups. The mice were euthanized 2 weeks after IH exposure, and the weight of tumor tissues was measured. Next, IL-6, IL-17, IL-10, and TNF-α levels in tumor tissues were measured via enzyme linked immunosorbent assay (ELISA), and hypoxia inducible factor-1α (HIF-1α) and transforming growth factor ß1 (TGF-ß1) expressions were examined through Western blot analysis. RESULTS: Two weeks of IH exposure significantly accelerated the growth of S180 solid tumors. Western blot analysis results showed that the expression levels of HIF-1α and TGF-ß1 in S180 tumors in the IH group were significantly upregulated compared with those in the RA group. ELISA results showed that compared with the RA group, the IH group had significantly increased TNF-α and IL-10 (P < 0.05) and significantly decreased IL-17 (P < 0.05). CONCLUSION: IH might promote the growth of S180 solid tumors by inhibiting the antitumor immune response and inducing tumor immune escape via the upregulation of TGF-ß1.

Synthesis of 99mTc-labeled 2-Mercaptobenzimidazole as a novel radiotracer to diagnose tumor hypoxia.

Discovery of 99mTc-labeled imidazole derivatives as a potential radiotracer for hypoxic tumor imaging is considered to be of great interest because of non-invasive detection capabilities. 2-Mercaptobenzimidazole (2-MBI) was successfully synthesized, characterized and radiolabeled with [99mTc (CO)3(H2O)3]+ intermediate to form 99mTc-2-MBI complex with radiochemical purity of ≥95% yield as observed by instant-thin layer chromatography (ITLC) and radio-high performance liquid chromatography (radio-HPLC). The 99mTc-2-MBI complex was observed to be stable in saline and serum with no noticeable decomposition at room temperature and 37 °C, respectively, over a time period of 24 h. Biodistribution results in Balb/c mice bearing S180 tumor show that 99mTc-2-MBI highly internalized in tumor tissue, also possess preferably high tumor/muscle and tumor/blood ratios 4.14 ±â€¯0.77 and 3.91 ±â€¯0.63, respectively at 24 h incubation. Scintigraphic imaging study shows 99mTc-2-MBI is visibly accumulated in hypoxic tumor tissue, suggesting it would be a promising radiotracer for early stage diagnosis of tumor hypoxia.

PGE2-HIF1α reciprocal induction regulates migration, phenotypic alteration and immunosuppressive capacity of macrophages in tumor microenvironment.

BACKGROUND: Very little is known about the role inflammation and mechanism(s) that enables the tumor to evade host's anti-tumor immune function during very initial days of tumor establishment. Our study focuses on the immune response and local inflammation specially the pro-inflammatory and immune modifier components that are responsible for tumor-induced immune-suppression, tumor-associated macrophages (TAM) at tumor microenvironment in mouse model from very early to late phase of tumor progression. METHODS: 1 × 105 Ascites tumor, EAC in Swiss albino or Sarcoma-180 (S-180) in Balb c mice strain were inoculated intra-peritonially and grouped into Control (0 day or no tumor), initial phase (3 day tumor), early (7 Day), Late (14 day) and terminal (21 day tumor) sets. T cell activity, tumor niche macrophage, inflammatory signatures were studied using Confocal microscopy, flowcytometry, ELISA, q-RT PCR and Western blot. RESULTS: We observed increased T cell infiltration at a very early stage of tumorigenesis in the tumor site with elevated percentage of activated/memory T cells. But increased cellular death and functional suppression of tumor site T cells during final stages. We observed increased infiltration of TAMs with skewed M2 phenotype. Increased chemokine receptor expression could be noted on these TAMs. Using HIF-1α inhibitor and prostaglandin receptor antagonists we demonstrated crucial role of these factor in functional alteration in TAMs. HIF-1α inhibition and also by prostaglandin receptor inhibition reduced signature pro-inflammatory gene expression, migration of macrophages and T cell suppression capacity of TAMs. We also demonstrated that PGE2 can induce HIF-1α activation in relatively less hypoxic microenvironment during early stages of tumor. CONCLUSION: Altogether, these findings strongly suggest link between prostaglandin mediated early HIF-1α activation and subsequent hypoxia induced HIF-1α activation that further enhances prostaglandin synthesis driving the recruitment and functional alteration of tumor site macrophages.

MAL adaptor (TIRAP) S180L polymorphism and severity of disease among tuberculosis patients.

OBJECTIVE: Though several genetic variants have been recognized to be associated with susceptibility to Tuberculosis (TB) infection and disease, a recent observation on the association of TIRAP C975T (S180L) variants with TB disease severity in mice model prompted us to assess their relevance in humans. In addition, TIRAP variants have also been reported to be associated with varied circulating Interferon-gamma induced protein (IP-10) levels. We investigated the association of TIRAP variants with severity of TB disease and IP-10 production in humans, which may be useful in predicting poor clinical outcome. METHODS: Culture positive symptomatic adult pulmonary TB (PTB) patients enrolled between August 2014 and October 2017 were included in this investigation. Allelic discrimination PCR and conventional IP-10 quantification methods were employed for genotyping and IP-10 measurement followed by statistical investigations to analyse patients' variables. RESULTS: Among 211 participants, C/C allele was identified in 70% (n = 147); 26% (n = 55) and 4% (n = 9) had C/T and T/T alleles respectively. There was no significant association between TIRAP variants and smear grade, chest-X-ray score, symptom severity score and circulating IP-10 levels. However, significant association was observed between i) circulating IP-10 levels and time to Mycobacterium Growth Indicator Tube (MGIT) culture conversion (p =0.032); ii) smear grade among active TB patients and circulating IP-10 levels (p =0 .032). CONCLUSIONS: Although mice experiments showed promising results with more severe disease in C/C and T/T individuals, we did not observe any such association in humans.

Low-dose paclitaxel combined with thalidomide inhibits angiogenesis in mice bearing S180 sarcoma / 中国肿瘤临床

Objective: To investigate the effect of low-dose paclitaxel combined with thalidomide on angiogenesis in mice bearing S180 sarcoma. Methods: An S180 sarcoma-bearing mouse model was established. Forty tumor-bearing mice were randomly divided into four groups: A, the control group, administered normal saline; B, the paclitaxel group, administered 20 mg/kg paclitaxel by intraperitoneal injection three times per week for 2 weeks; C, the thalidomide group, administered 200 mg/kg thalidomide by gavage three times per week for 2 weeks; and D, the paclitaxel + thalidomide group, administered paclitaxel (20 mg/kg) and thalidomide (200 mg/kg) by intraperitoneal/intragastric administration three times per week for 2 weeks. Tumor weight, tumor inhibition rate, microvascular density (MVD), and vascular endothelial growth factor (VEGF) expression were measured. Results: Tumor weight, VEGF expression, and MVD were lower in groups B, C, and D (P<0.05) than in the control. VEGF expression and MVD were lower in group D than in groups B and C; these differences were statistically significant (P<0.05). Conclusions: Low-dose paclitaxel combined with thalidomide exerted an inhibitory effect on vascular growth in S180 sarcoma.

The Synthesis and Biological Function of a Novel Sandwich-Type Complex Based on {SbW9 } and Flexible bpp Ligand.

A novel sandwich-type complex [Na(H2 O)4 ][{Na3 (H2 O)5 }{Mn3 (bpp)3 } (SbW9 O33 )2 }]·8H3 O (MnSbW-bpp) (bpp = 1,3-bis(4-pyridyl) propane) is synthesized and characterized by elemental analysis, IR, thermogravimetric analysis, and single-crystal X-ray diffraction. The MnSbW-bpp compound is the first sandwich case bridged by a flexible ligand. Its biological function of MnSbW-bpp in antitumor activity is also determined in vitro and in vivo. The inhibitory proliferation and induction of apoptosis are performed by flow cytometry assay, S180 (sarcoma) tumor xenograft in ICR mice, the color Doppler ultrasound monitor, and TdT-mediated dUTP-biotin nick end labeling assay. The results show that the novel compound-MnSbW-bpp-is synthesized and identified by its physical and chemical characteristics, such as the fluorescent and paramagnetic activities. MnSbW-bpp indicates a potency inhibition of human cancer lines, such as SGC-7901, HT-29, HepG2, Hela, U2OS, SaoS2, and HMC cells. MnSbW-bpp also inhibits the growth of tumor xenograft in mice, induced cell apoptosis, and released cytochrome c in vivo and in vitro. Thus, MnSbW-bpp, as a new compound, possesses the potent inhibition of cancer cells, which indicates that the MnSbW-bpp has potential merit for the further evaluation of a novel antitumor agent.

Ascorbic acid and retinol palmitate modulatory effect on omeprazole-induced oxidative damage, and the cytogenetic changes in S. cerevisiae and S180 cells.

Omeprazole (OM), a prototype proton pump inhibitor, oxidizes thiol groups and induces DNA damage. The aim of this study was to evaluate the oxidative effects of omeprazole and its interactions with ascorbic acid (AA, 50 µM) and retinol palmitate (RP) in proficient and deficient Saccharomyces cerevisiae strains, as well as levels of cytogenetic damage in Sarcoma 180 (S180) cells. Omeprazole was tested at concentrations of 10, 20 and 40 µg/mL, whereas H2O2 (10 mM), cyclophosphamide (20 mg/mL), and saline (0.9% NaCl solution) were employed as stressor, positive control, and negative control, respectively. Results revealed that omeprazole concentration-dependently induces oxidative effects in S. cerevisiae strains. However, omeprazole co-treated with ascorbic acid (50 µM) and retinol palmitate (100 IU) significantly modulated the oxidative damage inflected on the S. cerevisiae strains. Furthermore, omeprazole did not produce micronucleus formation and chromosomal bridges in S180 cells, but induced shoots. Significant increase in karyolysis and karyorrhexis were also observed with the omeprazole treated groups, which was modulated by co-treatment with ascorbic acid and retinol palmitate. Taken all together, it is suggested that ascorbic acid and retinol palmitate can substantially modulate the oxidative damage caused by omeprazole on the S. cerevisiae strains, however, much precaution is recommended with omeprazole and antioxidant co-treatment.

Effective capture of circulating tumor cells from an S180-bearing mouse model using electrically charged magnetic nanoparticles.

BACKGROUND: Technology enabling the separation of rare circulating tumor cells (CTCs) provides the potential to enhance our knowledge of cancer metastasis and improve the care of cancer patients. Modern detection approaches commonly depend on tumor antigens or the physical size of CTCs. However, few studies report the detection of CTCs by the electrical differences between cancer cells and normal cells. RESULTS: In this study, we report a procedure for capturing CTCs from blood samples using electrically charged superparamagnetic nanoparticles (NPs). We found that only positively charged NPs attached to cancer cells, while negatively charged NPs did not. The capture method with positively charged NPs offered a sensitivity of down to 4 CTCs in 1 mL blood samples and achieved a superior capture yield (> 70%) for a high number of CTCs in blood samples (103-106 cells/mL). Following an in vitro evaluation, S180-bearing mice were employed as an in vivo model to assess the specificity and sensitivity of the capture procedure. The number of CTCs in blood from tumor-bearing mice was significantly higher than that in blood from healthy controls (on average, 75.8 ± 16.4 vs. zero CTCs/100 µL of blood, p < 0.0001), suggesting the high sensitivity and specificity of our method. CONCLUSIONS: Positively charged NPs combined with an in vivo tumor model demonstrated that CTCs can be distinguished and isolated from other blood cells based on their electrical properties.

An-tumor activity of sesquiterpenoids from ginseng in S180 tumor-bearing mice and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the anti-tumor activity in vivo of sesquiterpenoids from ginseng (SPG) in the S180 tumor-bearing mice, and to clarify its mechanism Methods: The ICR male mice were used to establish the S180 tumor-bearing models. The model mice were randomly divided into model group, 5-fluorouracil (5-FU) group (20 mg • k g- 1 , ip), low dose of SPG group (2. 5 mg • k g- 1 , ig), and high dose of SPG group (10. 0 mg • k g- 1 , ig), 8 mice in each group; another 8 mice were selected and used as blank control group. Fourteen days after administration, the blood samples were collected from the eyes of all mice, and they were sacrificed by cervical dislocation, then the tumor tissue was excised and weighed; the inhibitory rate of tumor was calculated; the serum levels of alanine aminotransferase (AST), aspartate aminotransferase (ALT), urea nitrogen (BUN), interleukin-2 (IL-2), tumor necrosis factor-a (TNF-a) and vascular endothelial growth factor (VEGF) of the mice in various groups were measured; the pathological changes of the tumor tissue of the mice in various groups were observed by HE staining; the expression levels of anti-apoptotic factor Bel-2, VEGF, p38 against mitogen-activated protein kinase (p38MAPK) and p-p38 against mitogen-activated protein kinase (p-p38MAPK) in the tumor tissue of the mice in various groups were analyzed by Western blotting method. Results: The inhibitory rates of the mice in SPG groups were increased significantly as the increase of SPG dose, and the inhibitory rate of tumor of the mice in high dose of SPG group was 76. 29%. Compared with model group, the serum levels of IL-2 and TNF-a of the mice in SPG groups were significantly increased (P < 0 . 01) and the levels of ALT, AST, BUN and VEGF were significantly decreased (P < 0. 05 or P< 0. 01). The HE staining results showed that the cells in model group were arranged neatly, a large number of nuclei were observed and the growth state was good; in low and high doses of SPG group, the number of nuclei in the tumor tissue of the mice was significantly reduced, the arrangement was loose, and a large area of necrosis occurred. The Western blotting results showed that compared with model group, the expression levels of Bcl-2 and VEGF proteins were significantly decreased (P < 0 . 01) and the expression levels of p-p38MAPK protein in low and high doses of SPG groups were significantly increased (P < 0.01). Conclusion: SPG has a good anti-tumor effect in the S180 tumor-bearing mice, and its mechanism may be associated with the decreasing of Bcl-2 and VEGF expressions and activation of p38 MAPK protein channel.

Isoliquiritigenin inhibits mouse S180 tumors with a new mechanism that regulates autophagy by GSK-3ß/TNF-α pathway.

In this study, the pharmacokinetic properties and stability of isoliquiritigenin (ILQ) in microsomes were evaluated. The data showed ILQ administrated by i.h had high absorption degree (absolute bioavailability> 90%), and strong elimination ability (average t1/2≈ 67 min). ILQ in rat tissues could reach peak at 0.25 h, and be detected in almost all tissues. In vitro, stability of ILQ in four species liver microsomes were rat > beagle dog > monkey > human > mouse. On the basis of pharmacokinetic (PK) profiles, mechanism of ILQ against S180 was explored. ILQ could not inhibit S180 growth directly in vitro. However, ILQ extremely prohibited S180 tumor volume in vivo. And when TNF-α in NK cells was knocked down by siRNA, ILQ had no inhibiting effect on S180 tumor. ILQ enhanced TNF-α expression in NK cells by FCM detection. Autophagy-associated proteins LC3-II, Beclin-1, ATG-7 were elevated in S180 cells co-cultured with ILQ treating NK cells. When TNF-α was knocked down by siRNA, ILQ could not induce autophagy in S180 tumors. In the NK cells of osteosarcoma patients, TNF-α was negatively correlated with GSK-3ß by ELISA detection. ILQ could inhibit GSK-3ß expression and further increased p65 and c-Rel expression in NK cells. When GSK-3ß was knocked down by siRNA, ILQ did not affect p65 and c-Rel expression. ILQ directly inhibited GSK-3ß and then activated the NF-κB pathway to enhance TNF-α expression in NK cells, which could induce autophagy in sarcomas. The present study supplied a new mechanism for ILQ against tumors.

Saponin from Tupistra chinensis Bak Inhibits NF-κB Signaling in Sarcoma S-180 Cell Mouse Xenografts.

This study examined the effect of saponins from Tupistra chinensis Bak (STCB) on the growth of sarcoma S-180 cells in vitro and in mouse xenografts as well as the underlying mechanisms. Cell proliferation was assessed by MTT assay. Cell cycle distribution was determined by flow cytometry. Sarcoma S-180 tumor-bearing mice were treated with different doses of STCB with 10 µg/mL 5-fluorouracil (5-Fu) as a positive control. The activity of nuclear factor (NF)-κB was detected by gel mobility shift assay. The mRNA level of NF-κB was determined by real-time quantitative RT-PCR. The results showed that in vitro STCB inhibited the growth of S-180 cells in a concentration-dependent manner, which was accompanied by cell cycle arrest at S-phase. In vivo STCB significantly inhibited the growth of S-180 tumor mouse xenografts in a dose-dependent manner with apparent induction of cell apoptosis. Moreover, STCB inhibited the activity of NF-κB p65 and reduced the expression of NF-κB p65 mRNA in mouse xenografts. It was concluded that STCB inhibits the proliferation and cell cycle progression of S-180 cells by suppressing NF-κB signaling in mouse xenografts. Our findings suggest STCB is a promising agent for the treatment of sarcoma.

Novel combination of 2-methoxyestradiol and cyclophosphamide enhances the antineoplastic and pro-apoptotic effects on S-180 ascitic tumour cells.

Sarcoma 180 (S-180) tumour cell line is a stable murine tumour cell line with 98-99% stumour takes capacity in Swiss albino mouse - Mus musculus. 2 Methoxyestradiol (2ME) - a promising anti-neoplastic and anti-angiogenic agent, showed toxicity to host body in higher concentration. Cyclophosphamide (CP), the anti-neoplastic agent has long been used as a chemotherapeutic drug for treatment of different cancers. Our studies have shown that the combination effect of 2ME and CP on S-180 tumour cell line is anti-proliferative and less toxic. The treatment with lower concentrations of 2ME and CP (6.5 mg 2ME/kg body weight + 75 mg CP/kg body weight) antagonistically increased the life span of tumour bearing mice and synergistically inhibited the viable cell population. 2ME or CP treatment individually induces G2/M arrest. The combination treatment of 2ME + CP (6.5 mg 2ME/kg body weight + 75 mg CP/kg body weight) produced a significant increase of cells in the G0 which is the indication of cell arrest or apoptosis. Reduction of cell viability by 2ME + CP treatments is due to apoptotic cell death. This combination therapy produced a significant inhibitory effect of cell proliferation and augmentation of cell accumulation in the G0 phase (i.e. apoptosis). Apoptosis is validated by Fluorescence staining of control and treated S-180 tumour cells with Acridine Orange and EtBr dye. Moreover, a steady increase in the frequency of complex chromosomal aberrations (i.e. tri-, qudri-radial translocations) in tumour cells was noted in that particular concentration of combination therapy treated series along with the increase in dead cell frequency and tumour regression pattern. It is assumed that, these chromosomal abnormalities or damages recorded in higher frequency prevent the affected metaphases to enter into the next cell cycle through apoptosis or necrosis. This study introduces a novel combination, where this particular concentration of 2ME + CP (i.e. 6.5 mg 2ME/kg body weight + 75 mg CP/kg body weight) not only enhanced the life span of tumour bearing mouse and decreased the tumour volume antagonistically but also inhibited the viable cell population synergistically, which could serve as a potential effective regimen for cancer treatment.

Saponin from Tupistra chinensis Bak Inhibits NF-κB Signaling in Sarcoma S-180 Cell Mouse Xenografts / 华中科技大学学报（医学）（英德文版）

This study examined the effect of saponins from Tupistra chinensis Bak (STCB) on the growth of sarcoma S-180 cells in vitro and in mouse xenografts as well as the underlying mechanisms.Cell proliferation was assessed by MTT assay.Cell cycle distribution was determined by flow cytometry.Sarcoma S-180 tumor-bearing mice were treated with different doses of STCB with 10 μg/mL 5-fluorouracil (5-Fu) as a positive control.The activity of nuclear factor (NF)-κB was detected by gel mobility shift assay.The mRNA level of NF-κB was determined by real-time quantitative RT-PCR.The results showed that in vitro STCB inhibited the growth of S-18 0 cells in a concentration-dependent manner,which was accompanied by cell cycle arrest at S-phase.In vivo STCB significantly inhibited the growth of S-180 tumor mouse xenografts in a dose-dependent manner with apparent induction of cell apoptosis.Moreover,STCB inhibited the activity of NF-κB p65 and reduced the expression of NF-κB p65 mRNA in mouse xenografts.It was concluded that STCB inhibits the proliferation and cell cycle progression of S-180 cells by suppressing NF-κB signaling in mouse xenografts.Our findings suggest STCB is a promising agent for the treatment of sarcoma.

Saponin from Tupistra chinensis Bak Inhibits NF-κB Signaling in Sarcoma S-180 Cell Mouse Xenografts / 华中科技大学学报（医学）（英德文版）

This study examined the effect of saponins from Tupistra chinensis Bak (STCB) on the growth of sarcoma S-180 cells in vitro and in mouse xenografts as well as the underlying mechanisms.Cell proliferation was assessed by MTT assay.Cell cycle distribution was determined by flow cytometry.Sarcoma S-180 tumor-bearing mice were treated with different doses of STCB with 10 μg/mL 5-fluorouracil (5-Fu) as a positive control.The activity of nuclear factor (NF)-κB was detected by gel mobility shift assay.The mRNA level of NF-κB was determined by real-time quantitative RT-PCR.The results showed that in vitro STCB inhibited the growth of S-18 0 cells in a concentration-dependent manner,which was accompanied by cell cycle arrest at S-phase.In vivo STCB significantly inhibited the growth of S-180 tumor mouse xenografts in a dose-dependent manner with apparent induction of cell apoptosis.Moreover,STCB inhibited the activity of NF-κB p65 and reduced the expression of NF-κB p65 mRNA in mouse xenografts.It was concluded that STCB inhibits the proliferation and cell cycle progression of S-180 cells by suppressing NF-κB signaling in mouse xenografts.Our findings suggest STCB is a promising agent for the treatment of sarcoma.

Antitumor and Immunoregulatory Activities of Seleno-ß-Lactoglobulin on S180 Tumor-Bearing Mice.

Degeneration of immune organs like thymus and spleen has been discovered in tumor-bearing mice; which increases the difficulties on oncotherapy. More effective drugs which target the protection of immune organs are expected to be researched. In this study; we aim to analyze the antitumor and immunoregulatory activities of seleno-ß-lactoglobulin (Se-ß-lg) on S180 tumor-bearing mice. Results indicated that Se-ß-lg exhibited a remarkable inhibitory effect on S180 solid tumors with the inhibition rate of 48.38%; and protected the thymuses and spleens of S180-bearing mice. In addition, Se-ß-lg could also balance the proportions of CD4⁺ and CD8⁺ T cells in spleens; thymuses and peripheral bloods; and improve Levels of IL-2; IFN-γ; TNF-α in mice serums. ß-lg showed weaker bioactivities while SeO2 showed stronger toxicity on mice. Therefore our results demonstrated that Se-ß-lg possessed stronger antitumor and immunoregulatory activities with lower side effects and had the potential to be a novel immunopotentiator and antitumor agent.