Tumor cell lysate with low content of HMGB1 enhances immune response of dendritic cells against lung cancer in mice / 南方医科大学学报

OBJECTIVE@#To assess the effect of tumor cell lysate (TCL) with low high-mobility group B1 (HMGB1) content for enhancing immune responses of dendritic cells (DCs) against lung cancer.@*METHODS@#TCLs with low HMGB1 content (LH-TCL) and normal HMGB1 content (NH-TCL) were prepared using Lewis lung cancer (LLC) cells in which HMGB1 was inhibited with 30 nmol/L glycyrrhizic acid (GA) and using LLC cells without GA treatment, respectively. Cultured mouse DCs were exposed to different doses of NH-TCL and LH-TCL, using PBS as the control. Flow cytometry was used to detect the expressions of CD11b, CD11c and CD86 and apoptosis of the stimulated DCs, and IL-12 levels in the cell cultures were detected by ELISA. Mouse spleen cells were co-cultured with the stimulated DCs, and the activation of the spleen cells was assessed by detecting CD69 expression using flow cytometry; TNF-β production in the spleen cells was detected with ELISA. The spleen cells were then co-cultured with LLC cells at the effector: target ratios of 5:1, 10:1 and 20:1 to observe the tumor cell killing. In the animal experiment, C57/BL6 mouse models bearing subcutaneous LLC xenograft received multiple injections with the stimulated DCs, and the tumor growth was observed.@*RESULTS@#The content of HMGB1 in the TCL prepared using GA-treated LLC cells was significantly reduced (P < 0.01). Compared with NH-TCL, LH-TCL showed a stronger ability to reduce apoptosis (P < 0.001) and promote activation and IL- 12 production in the DCs. Compared with those with NH-TCL stimulation, the DCs stimulated with LH-TCL more effectively induced activation of splenic lymphocytes and enhanced their anti-tumor immunity (P < 0.05). In the cell co-cultures, the spleen lymphocytes activated by LH-TCL-stimulated DCs showed significantly enhanced LLC cell killing activity (P < 0.01). In the tumor-bearing mice, injections of LH-TCL-stimulated DCs effectively activated host anti-tumor immunity and inhibited the growth of the tumor xenografts (P < 0.05).@*CONCLUSION@#Stimulation of the DCs with LH-TCL enhances the anti-tumor immune activity of the DCs and improve the efficacy of DCbased immunotherapy for LLC in mice.

Circular RNA hsa_circ_0000317 inhibits non-small cell lung cancer progression through regulating microRNA-494-3p/phosphatase and tensin homolog deleted on chromosome 10 axis

Abstract Background: Circular RNA (circRNA), a group of non-coding RNA, is pivotal in the progression of various cancers, including Non-Small Cell Lung Cancer (NSCLC). Some circRNAs have been reported to be implicated in the progression of NSCLC, however, the function and molecular mechanism of hsa_circ_0000317 (circ_0000317) in NSCLC have not been fully understood. Methods: The significantly differentially expressed circRNA in NSCLC tissues, circ_0000317, was screened out by microarray. Circ_0000317, microRNA(miR)-494-3p and Phosphatase and Tensin Homolog Deleted on Chromosome 10 (PTEN) expressions in NSCLC tissues were respectively probed by quantitative real-time polymerase chain reaction and western blot assay. MTT and Transwell assays were adopted to examine the growth, migration, and invasion of NSCLC cells. Bioinformatics, luciferase reporter gene assay, RNA immunoprecipitation, and RNA pull-down assay were conducted to probe the relationships among circ_0000317, miR-494-3p, and PTEN. Results: Circ_0000317 expression level was reduced in NSCLC tissues and cell lines. Circ_0000317 expression in NSCLC patients was associated with TNM stage and lymphatic metastasis. Circ_0000317 overexpression restrained the proliferation, migration, and invasion of NSCLC cells, but co-transfection of miR-494-3p mimics partially reversed this effect. In addition, circ_0000317, was identified as a competitive endogenous RNA, which could sponge miR-494-3p to increase PTEN expression and activate PI3K/AKT pathway. Conclusion: Circ_0000317, inhibits NSCLC progression via modulating miR-494-3p/PTEN/PI3K/AKT pathway.

Effect and Mechanism of Berberine Hydrochloride Against Lung Cancer Cells in Vitro Based on Mevalonate Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the efficacy and mechanism of berberine hydrochloride （BBH） against lung cancer cells through the mevalonate （MVA） pathway. MethodHuman lung cancer A549 cells and mouse Lewis lung carcinoma （LLC） cells were used as research subjects. Cell proliferation and cell counting kit-8 （CCK-8） assay were performed to detect the inhibitory effect of BBH （10， 20， 30， 40， 50 μmol·L-1） on the proliferation of the two kinds of cells （48 h）. Then cell scratch assay was used to explore the influence of BBH （40 μmol·L-1） on the migration of A549 and LLC cells （24， 48 h）， and colony formation assay was conducted to compare the colony formation ability of the cells under different concentrations of BBH （10， 20， 40 μmol·L-1）. Moreover， the effects of BBH （40 μmol·L-1） on the content of acetyl-coenzyme A （A-CoA） and total cholesterol （TC） in A549 and LLC cells were determined by kit assay. AutoDock Vina was used for the dock of BBH and MVA pathway regulatory protein， sterol regulatory element-binding protein 2 （SREBP2）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to observe the effects of BBH （40 μmol·L-1） on the mRNA expression of nine genes related to the MVA pathway in A549 and LLC cells： hydroxymethylglutaryl-CoA synthase 1 （HMGCS1）， hydroxymethylglutaryl-CoA Reductase （HMGCR）， mevalonate kinase （MVK）， phosphomevalonate kinase （PMVK）， mevalonate 5-pyrophosphate decarboxylase （MVD）， farnesyl diphosphate synthase （FDPS）， squalene epoxidase （SQLE）， farnesyl-diphosphate farnesyltransferase 1 （FDFT1）， and geranylgeranyl diphosphate synthase 1 （GGPS1）. Western blot was performed to clarify the effects of BBH （40 μmol·L-1） on the expression of three key proteins of the MVA pathway： HMGCS1， HMGCR， and FDFT1. The Cancer Genome Atlas （TCGA） database was searched to analyze the relationship between HMGCS1， HMGCR， FDFT1 and transcription gene SREBF2 in non-small cell lung cancer （NSCLC）. ResultCompared with the conditions in the control group， the proliferation， migration， and colony formation of A549 and LLC cells in the BBH group were decreased （P<0.01）， while the cell apoptosis rate was increased （P<0.01）. Molecular docking showed that BBH had good binding activity with SREBP2. In addition， the content of A-CoA and TC of the MVA pathway was reduced （P<0.01）. BBH down-regulated the mRNA expression of HMGCS1， HMGCR， MVK， PMVK， MVD， FDPS， SQLE， FDFT1， and GGPS1 in A549 and LLC cells （P<0.01）， and lowered the levels of HMGCS1， HMGCR， and FDFT1 proteins （P<0.05， P<0.01）. In NSCLC patients， HMGCS1， HMGCR， and FDFT1 were highly correlated with SREBF2 （R=0.54， R=0.57， and R=0.48）. ConclusionBBH can inhibit the proliferation， migration， and colony formation of A549 and LLC cells and promote cell apoptosis， which may be related to the regulation of MVA pathway by BBH binding to SREBP2.

Effect and mechanism of aspirin combined with vinorelbine on non-small cell lung cancer / 中国中药杂志

The study aimed to investigate the effect and mechanism of aspirin combined with vinorelbine on the proliferation and apoptosis of non-small cell lung cancer cells. 3-(4-dimethylthiazolyl-2)-2-diphenyltetrazolium bromide(MTT) was used to detect the cytotoxic effect of aspirin and vinorelbine on H460 and A549 cells, and half of inhibitory concentration(IC_(50)) value of drugs as well as synergistic effect were calculated. The results showed that both aspirin and vinorelbine inhibited the cancer cells proliferation by a concentration-dependent manner with IC_(50 )values of 1.553 mmol·L~(-1) and 0.033 μmol·L~(-1) in H460 cells, respectively. The IC_(50 )values of aspirin and vinorelbine were 1.70 mmol·L~(-1)and more than 20 μmol·L~(-1) in A549 cells. The combination index(CI) value was used to evaluate the combined effect of two drugs. Aspirin combined with vinorelbine had synergistic effects at the ratio of 100∶1 on H460 cells and 1∶10 on A549 cells(CI<1). Clone formation and 4',6-diamidino-2-phenylindole(DAPI)/propidium iodide(PI) staining assays were used to verify the effect of the combination of two drugs on proliferation of H460 cells. Compared with the aspirin single group, the combination group had stronger inhibitory effect on the proliferation of H460 cells and the clone formation rate was 49.5%(P<0.05). Furthermore, apoptosis, mitochondrial membrane potential, reactive oxygen species and Western blot experiments were used to explore the synergistic mechanism of aspirin combined with vinorelbine in inhibiting cell proliferation. The results showed that the cancer cell apoptosis rate was 52.8%, the mitochondrial membrane potential was decreased to 33.1%, and the levels of reactive oxygen species was increased to 73.3% in combination group, which were significantly different from those of the single drug treatment groups(P<0.05). Western blot showed that combination group significantly up-regulated the expressions of Bax, p53, cleaved caspase-3 and cytochrome C, while down-regulated the expression of anti-apoptosis proteins such as Bcl-xL and Bcl-2 when compared with single groups. Our results suggested that aspirin combined with vinorelbine could synergistically inhibit the proliferation of H460 cells by inducing the cell apoptosis through the mitochondrial pathway.

Effect of Huaier aqueous extract on growth and metastasis of human non-small cell lung cancer NCI-H1299 cells and its underlying mechanisms / 中国中药杂志

This study aims to investigate the effect of Huaier aqueous extract on the growth and metastasis of human non-small cell lung cancer NCI-H1299 cells and its underlying mechanisms. MTT assay was used to detect the effect of Huaier aqueous extract on the proliferation of NCI-H1299 cells. Flow cytometry was used to examine the effect of Huaier aqueous extract on the apoptosis, cell cycle, and ROS level of NCI-H1299 cells. Wound healing assay was used to evaluate the effect of Huaier aqueous extract on the migration ability of NCI-H1299 cells. Western blot was used to detect the levels of proteins involving apoptosis, epithelial-mesenchymal transition(EMT), and MAPK signaling pathway in NCI-H1299 cells exposed to Huaier aqueous extract. The results showed that Huaier aqueous extract inhibited the proliferation of NCI-H1299 cells, and induced cell-cycle arrest at the phase S. Huaier aqueous extract promoted the apoptosis of NCI-H1299 cells by down-regulating the expression of anti-apoptotic protein Bcl-2. Moreover, Huaier aqueous extract increased ROS level and induced ferroptosis in NCI-H1299 cells. EMT played a critical role in cancer metastasis. Huaier aqueous extract reduced the migration ability of NCI-H1299 cells by inhibiting EMT of NCI-H1299 cells. In addition, this study revealed that Huaier aqueous extract inhibited MAPK signaling pathway in human non-small cell lung cancer NCI-H1299 cells, which may be one of Huaier's mechanisms in inhibiting growth and metastasis of NCI-H1299 cells. This study provides a new theoretical basis for the clinical treatment of lung cancer with Huaier, and important reference significance for further studies on the anti-tumor mechanisms of Huaier.

Yiqi Huoxue decoction inhibits malignant biological behavior of human lung cancer cells via the miR-21/PTEN signaling pathway / 中国临床药理学与治疗学

AIM: To investigate the inhibitory effect of Yiqi Huoxue decoction on the malignant biological behavior of lung cancer cells and its mechanism. METHODS: Human lung cancer A549 cells were treated with different doses of Yiqi Huoxue Decoction serum (5%, 10%, 15%). CCK-8 assay, transwell chamber experiment, flow cytometry, Western blot and qRT-PCR method were used to study the effect of different doses of Yiqi Huoxue Decoction-containing serum to cell proliferation, cell migration and invasion, cell apoptosis, PTEN protein and miR-21 expression. RESULTS:Compared with the drug-free serum group, survival rate, migration and invasion ability of A549 cells decreased after treatment with different doses of drug-containing serum. The apoptosis rate of A549 cells increased, PTEN mRNA and the expression of its protein increased, the expression of miR-21 decreased, and the medium-dose (10%) drug-containing serum group had the best effect. After the transfection of miR-21 mimics, miR-21 expression was up-regulated, while PTEN protein expression was down-regulated in cells. PTEN protein expression was up-regulated after treatment with medium-dose (10%) drug-containing serum. CONCLUSION: Yiqi Huoxue Decoction can effectively inhibit the malignant cell biological behavior of human lung cancer A549 cells and may be related to the regulation of the miR-21/PTEN signaling pathway.

Quantitative proteomics of inhibitory mechanism of Dendrobium denneanum ether extract on lung cancer cells / 中国中药杂志

Dendrobium denneanum have been used for a long time as rare medicinal herbs in traditional Chinese medicine. Our previous works found that ether extract of D. denneanum had higher anticancer activities than alcohol or water extract,thus with better development prospects. Quantitative proteomics based on SILAC technique was used to investigate the anticancer mechanism of D. denneanum on lung tumor cell line A549,and 4 855 proteins were detected in A549 cells. Quantitative proteomics experiments found that 193 proteins of A549 cells were up-regulated,and 44 proteins were down-regulated by ether extract of D. denneanum. Those proteins are associated with synthesis,transport and metabolism of biological macromolecules,chaperone,DNA repair,oxidoreductase,cell adhesion,cell cycle,apoptosis and autophagy. Through the function analysis of differentially expressed proteins,it was inferred that ether extract of D. denneanum caused cell protein metabolism disorder,endoplasmic reticulum stress response,abnormal self-repair mechanism of cells,damage of cell adhesion and proliferation; besides,it caused a dramatic increase in ROS level in A549 cells,and upset the balance of intracellular oxidation reduction system. Affected by the above factors,lung cancer cells initiated apoptosis and autophagy,which accelerated cell death. This research explains the anticancer mechanism of D. denneanum from the perspective of quantitative proteomics,and lays a foundation for future research and development of new anticancer drugs based on ether extract of D. denneanum.

Screening of nucleic acid aptamer of lung cancer cells based on cell exponential enrichment ligand system evolution and its application in tumor diagnosis and treatment / 生物医学工程学杂志

Nucleic acid aptamer is an oligonucleotide sequence screened by the exponential enrichment ligand system evolution technology (SELEX). Previous studies have shown that nucleic acid aptamer has a good application prospect in tumor diagnosis and treatment. Therefore, we reviewed the selection and identification of nucleic acid aptamer of lung cancer cells in recent years, and discussed the effect of aptamer as targeting drugs and targeting vectors on the diagnosis of tumors, which provide a new idea for early diagnosis and treatment of tumor.

Evaluation of cytotoxicity of stearoyl gemcitabine-loaded PLGA nanoparticles to Lewis lung cancer cells / 中国免疫学杂志

Objective:To investigate the anti-proliferation effect of 4-(N)-stearoyl gemcitabine-loaded poly(lactic-co-glycolic) acid nanoparticles(GemC18-PLGA-NPs) on Lewis lung cancer cells(LLC) in vitro.Methods: Lewis cells were incubated with GemC18-PLGA-NPs,free GemC18,gemcitabine HCl(GemHCl) or GemC18-free blank nanoparticles(PLGA-NPs) respectively and cell viability was determined using an MTT assay after 24,48 or 72 h of incubation.The apoptosis rate after 48 and 72 h of incubation were measured by flow cytometry.Results:GemC18-PLGA-NPs,GemC18,and GemHCl all significantly inhibited the growth of LLC cells, and the survival rate of GemHCl group was lowest,GemC18-PLGA-NPs group had the highest survival rate.The cell survival rate of GemC18-PLGA-NPs after 72 h was significantly higher than that of GemHCl (P<0.05) at the concentration of 1 μmol/L,indicating that it had a significant drug release effect.PLGA-NPs group produced trifle inhibition on the Lewis cells without correlation to time or concentration.Conclusion:GemC18-PLGA-NPs have significant anti-proliferation effect on mouse Lewis lung cancer cells in vitro.

TIPE2 sensitizes CDDP to human non-small cell lung cancer cell line NCI-H1975 by suppressing AP-1 / 中国免疫学杂志

Objective:To investigate the chemosensitivity of TIPE2 in enhancing non-small cell lung cancer cell line NCl-H1975 and its mechanisms.Methods:TIPE2 lentiviral vector was transfected into NCI-H1975.IC50 was measured by CCK-8 method after treated with CDDP.Apoptotic cells were detected by Annexin V/FITC and PI apoptosis detection kit.The expression of AP-1 and MDR-1 were measured using Western blot after TIPE2 transfected.The mRNA expression of IL-1,IL-6 and TNF-α were measured using Real-time PCR after TIPE2 transfected combined with CDDP administration.Results:(1) TIPE2 reduces the values of IC50 of NCI-H1975 cells for CDDP(P＜0.001).(2) TIPE2 increases the apoptosis rate of NCI-H1975 cells when treated with CDDP(P＜0.05).(3) TIPE2 significantly reduces the expression of AP-1 and MDR1 in NCI-H1975 cells when treated with CDDP.(4)TIPE2 reduces the mRNA expression of IL-1,IL-6 and TNF-α in NCI-H1975 cells when treated with CDDP(P＜0.01).Conclusion:TIPE2 may increase the chemosensitivity of non-small cell lung cancer cell line NCl-H1975 to CDDP by inhibiting AP-1 protein.

Effect of Olaparib on proliferation activity and radiosensitization of human non-small cell lung cancer cells / 国际生物医学工程杂志

Objective To investigate the effects of Olaparib on cell proliferation and radiosensitization of human non-small cell lung cancer cells.Methods Non-small cell lung H460 and H1299 cell lines were cultured in vitro and the cells in logarithmic growth phase were selected for experiments.MTT and colony formation assays were used to determine cell proliferation and radiosensitization,respectively.Single cell gel electrophoresis assay (comet assay) was used to detect irradiation-induced DNA damage.Results The results of MTT assay showed that Olaparib inhibited the proliferation of H460 and H1299 cells in a dose-dependent pattern (all P＜0.05).H1299 cell line was more sensitive to Olaparib than H460 cells.The results of colony formation assay showed that Olaparib enhanced the radiosensitizition of H460 and H1299 cells (all P＜0.05).The results of comet assay showed that Olaparib increased γ ray-induced DNA damage.Conclusions Olapani can enhance the radiosensitization of human non-small cell lung cancer cells,and the radiosensitization effect of Olaparib may be associated with the inhibition of cell proliferation and induction of irradiation-induced DNA damage.

Antiproliferative effect of Momordica cochinchinensis seeds on human lung cancer cells and isolation of the major constituents

Abstract Gac, Momordica cochinchinensis (Lour.) Spreng., Cucurbitaceae, is an indigenous South Asian edible fruit and has been used therapeutically in Traditional Chinese Medicine. Previous studies have shown that M. cochinchinensis seed (Momordicae Semen) has various pharmaceutical properties such as antioxidant and anti-ulcer effects as well as contains secondary metabolites with potential anticancer activities such as triterpenoids and saponins. However, its biological activities in cancer have not yet been investigated. In this study, we found that its ethanol extract reduced cell proliferation in four human lung cancer cell lines, A549, H1264, H1299 and Calu-6. Phytochemical investigation of the ethanol extract was carried out, and resulted in isolation of two major saponins, which were identified as gypsogenin 3-O-β-d-galactopyranosyl(1 → 2)-[α-l-rhamnopyranosyl(1 → 3)]-β-d-glucuronopyranoside (1) and quillaic acid 3-O-β-d-galactopyranosyl(1 → 2)-[α-l-rhamnopyranosyl(1 → 3)]-β-d-glucuronopyranoside (2). Treatment with these isolated compounds (1 and 2) decreased cel1 proliferation in all human lung cancer cell lines tested. In addition, the compounds attenuated primary lung endothelial cell proliferation. Taken together, these findings suggest M. cochinchinensis seeds have antiproliferative activity on human lung cancer cells as well as angiostatic effect on lung endothelial cells.

Effects of Shanxian Granule on antagonistic immunity of Lewis lung cancer mice and level of IFN-γ,TNF-β,IL-10 in peripheral blood / 中国免疫学杂志

Objective:To investigate the effects of Shanxian granule on proliferation of Lewis lung cancer cells and anti-tumor immunity and immune microenvironment of Lewis lung cancer-bearing mice in order to explore the molecular mechanism of anti-tumor of Shanxian Granule and improve the anti-tumor immunity of the body, and provide further theoretical basis for its clinical application. Methods:Lewis lung cancer cells was transplanted to axillary skin to establish mouse tumor model. The mice divided into blank group,model group,chemotherapy group and Shanxian granule group. The tumor tissue of Lewis lung cancer tumor bearing mice was weighed and the tumor inhibition rate was calculated. Immunohistochemical method was used to detect the expression of CD and CD8 in spleen tissue. The effect of lymphocytes on the proliferation of Lewis lung cancer cells was detected by CCK-8 method. The level of IFN-γ,TNF-βand IL-10 in peripheral blood were detected by ELISA. Results:①The tumor inhibition rate of Lewis lung cancer was 45. 99% in Shanxian Granule group,which was significantly higher than that of chemotherapy group (P<0. 05).②The lymphocytes of mouse can inhibit the proliferation of Lewis lung cancer cells and have a positive correlation with lymphocyte concentration and duration of action. Moreover,CD4+ T cells,CD4+/CD8+ratio and lymphocyte inhibition rate of Lewis lung cancer cells in model group and chem-otherapy group were significantly lower than those in blank group (P<0. 05). Shanxian granule group was significantly higher than the model group and chemotherapy group ( P<0. 05 ) . However, there was no significant difference between Shanxian granule group and blank group(P>0. 05).③The levels of IFN-γand TNF-βin peripheral blood of model group and chemotherapy group were significantly lower than those in blank group,while IL-10 was significantly higher than that in blank group (P<0. 05). The levels of IFN-γand TNF-βin peripheral blood of mice in Shanxian granule group were significantly higher than those in model group and chemotherapy group, while IL-10 was significantly lower than that in model group and chemotherapy group (P<0. 05). There was no significant difference in IFN-γ,TNF-β and IL-10 in peripheral blood of mice between Shanxian granule group and blank group. Conclusion:Shanxian granule can significantly inhibit the growth of tumor tissue of Lewis lung cancer tumor bearing mice,increase the spleen index of mice,enhance the activity of T lymphocytes,upregulate IFN-γ and TNF-β in peripheral blood and decrease IL-I. These suggested that the anti-tumor effect of Shanxian granule may be achieved by regulating the content of CD4+ T lymphocyte,the ration of CD4+/CD8+ and Th1/Th2 ratio,in order to restore the immune steady function of tumor patients,improve the immune system and enhance the immune surveillance function.

Effects of Shanxian Granule on antagonistic immunity of Lewis lung cancer mice and level of IFN-γ,TNF-β,IL-10 in peripheral blood / 中国免疫学杂志

Objective:To investigate the effects of Shanxian granule on proliferation of Lewis lung cancer cells and anti-tumor immunity and immune microenvironment of Lewis lung cancer-bearing mice in order to explore the molecular mechanism of anti-tumor of Shanxian Granule and improve the anti-tumor immunity of the body, and provide further theoretical basis for its clinical application. Methods:Lewis lung cancer cells was transplanted to axillary skin to establish mouse tumor model. The mice divided into blank group,model group,chemotherapy group and Shanxian granule group. The tumor tissue of Lewis lung cancer tumor bearing mice was weighed and the tumor inhibition rate was calculated. Immunohistochemical method was used to detect the expression of CD and CD8 in spleen tissue. The effect of lymphocytes on the proliferation of Lewis lung cancer cells was detected by CCK-8 method. The level of IFN-γ,TNF-βand IL-10 in peripheral blood were detected by ELISA. Results:①The tumor inhibition rate of Lewis lung cancer was 45. 99% in Shanxian Granule group,which was significantly higher than that of chemotherapy group (P<0. 05).②The lymphocytes of mouse can inhibit the proliferation of Lewis lung cancer cells and have a positive correlation with lymphocyte concentration and duration of action. Moreover,CD4+ T cells,CD4+/CD8+ratio and lymphocyte inhibition rate of Lewis lung cancer cells in model group and chem-otherapy group were significantly lower than those in blank group (P<0. 05). Shanxian granule group was significantly higher than the model group and chemotherapy group ( P<0. 05 ) . However, there was no significant difference between Shanxian granule group and blank group(P>0. 05).③The levels of IFN-γand TNF-βin peripheral blood of model group and chemotherapy group were significantly lower than those in blank group,while IL-10 was significantly higher than that in blank group (P<0. 05). The levels of IFN-γand TNF-βin peripheral blood of mice in Shanxian granule group were significantly higher than those in model group and chemotherapy group, while IL-10 was significantly lower than that in model group and chemotherapy group (P<0. 05). There was no significant difference in IFN-γ,TNF-β and IL-10 in peripheral blood of mice between Shanxian granule group and blank group. Conclusion:Shanxian granule can significantly inhibit the growth of tumor tissue of Lewis lung cancer tumor bearing mice,increase the spleen index of mice,enhance the activity of T lymphocytes,upregulate IFN-γ and TNF-β in peripheral blood and decrease IL-I. These suggested that the anti-tumor effect of Shanxian granule may be achieved by regulating the content of CD4+ T lymphocyte,the ration of CD4+/CD8+ and Th1/Th2 ratio,in order to restore the immune steady function of tumor patients,improve the immune system and enhance the immune surveillance function.

The effect of c-IAP1 on radiosensitivty of lung cancer cells / 国际生物医学工程杂志

Objective To investigate the correlation between the level of apoptosis protein 1 (c-IAP1) and the radiosensitivity of lung cancer cells.Method The survival rate and proliferation of the lung cancer cells lines (A549,H460,H1299,H358,HCC827,H1650) from six human were detected by thiazolyl blue tetrazolium bromide (MTT) and cell colony formation assay.The DNA damage effects of radiation on lung cancer cells were detected by comet assay.The expressions of c-IAP1 protein and its mRNA were determined by Western blot and real-time quantitative PCR.Results The results of MTT and colony formation showed that the radiosensitivity of different lung cancer cells was also different,among which H358 and H460 cells had the highest radiosensitivity than that of H1650 and HCC827 cells,and H1299 and A549 cells had the weakest radiosensitivity.The results of comet assay showed that six kinds of lung cancer cells were suffered by DNA damage after radiation,and the DNA damage of H358 cells was most serious.The results of Western blot and real-time quantitative PCR showed that the c-IAP1 protein level was negatively correlated with the radiosensitivity of lung cancer cells.The higher the c-IAP1 protein level,the weaker the radiosensitivity of cells.The radiosensitivity was also affected by Smac protein levels.Conclusions c-IAP1 may be a selective target gene in mediating the radiosensitivity of lung cancer cells and this paper may contribute to the study of radioresistance and radiosensitization of cancer cell.

The effect of c-IAP1 on radiosensitivty of lung cancer cells / 国际生物医学工程杂志

Objective To investigate the correlation between the level of apoptosis protein 1 (c-IAP1) and the radiosensitivity of lung cancer cells.Method The survival rate and proliferation of the lung cancer cells lines (A549,H460,H1299,H358,HCC827,H1650) from six human were detected by thiazolyl blue tetrazolium bromide (MTT) and cell colony formation assay.The DNA damage effects of radiation on lung cancer cells were detected by comet assay.The expressions of c-IAP1 protein and its mRNA were determined by Western blot and real-time quantitative PCR.Results The results of MTT and colony formation showed that the radiosensitivity of different lung cancer cells was also different,among which H358 and H460 cells had the highest radiosensitivity than that of H1650 and HCC827 cells,and H1299 and A549 cells had the weakest radiosensitivity.The results of comet assay showed that six kinds of lung cancer cells were suffered by DNA damage after radiation,and the DNA damage of H358 cells was most serious.The results of Western blot and real-time quantitative PCR showed that the c-IAP1 protein level was negatively correlated with the radiosensitivity of lung cancer cells.The higher the c-IAP1 protein level,the weaker the radiosensitivity of cells.The radiosensitivity was also affected by Smac protein levels.Conclusions c-IAP1 may be a selective target gene in mediating the radiosensitivity of lung cancer cells and this paper may contribute to the study of radioresistance and radiosensitization of cancer cell.

Cytotoxic activity of Androctonus australis hector venom and its toxic fractions on human lung cancer cell line

Several studies have showed that animal venoms are a source of bioactive compounds that may inhibit the growth of cancer cells, which makes them useful agents for therapeutic applications. Recently, it was established that venom toxins from scorpions induced cytotoxic, antiproliferative and apoptogenic effects on cancer cells. Therefore, the present study aims to investigate the cytotoxic activity of Androctonus australis hector (Aah) scorpion venom and its toxic fractions (FtoxG-50 and F3) on NCI-H358 human lung cancer cells. Methods: The cytotoxic and antiproliferative activities were estimated using MTT assay, lactate dehydrogenase release and clonogenic assays. Apoptosis was evaluated by Hoechst 33258 staining, DNA fragmentation assay and caspase-3 activity. Oxidative stress was analyzed by reactive oxygen species, nitric oxide, malondialdehyde and protein carbonyl levels along with assessment of antioxidant status. In addition, alteration of mitochondrial membrane potential was analyzed by JC1 fluorescent dye. Results: The present findings showed that F3 fraction was more cytotoxic towards NCI-H358 lung cancer cells with an IC50 of 27.05 ± 0.70 g/mL than venom alone (396.60 ± 1.33 g/mL) and its toxic fraction FtoxG-50 (45.86 ± 0.91 g/mL). Nevertheless, F3 fraction was not cytotoxic at these concentrations on normal human lung fibroblast MRC-5 cells. Inhibition of NCI-H358 cell proliferation after F3 fraction exposure occurred mainly by apoptosis as evidenced by damaged nuclei, significant DNA fragmentation level and caspase-3 activation in a dose dependent manner. Moreover, F3 fraction enhanced oxidative and nitrosative stress biomarkers and dissipated mitochondrial membrane potential in lung cancer cells along with significant depletion in cellular enzymatic and non-enzymatic antioxidants. Further, the apoptosis induced by F3 fraction was markedly prevented by the antioxidant N-acetylcysteine (NAC) suggesting the potential mechanism of oxidative stress. Conclusion: These findings suggest that F3 fraction could induce apoptosis in lung cancer cells through involvement of oxidative stress and mitochondrial dysfunction. Hence, these properties make F3 fraction a promising candidate for development of new anticancer agents.

Cytotoxic activity of Androctonus australis hector venom and its toxic fractions on human lung cancer cell line

Background: Several studies have showed that animal venoms are a source of bioactive compounds that may inhibit the growth of cancer cells, which makes them useful agents for therapeutic applications. Recently, it was established that venom toxins from scorpions induced cytotoxic, antiproliferative and apoptogenic effects on cancer cells. Therefore, the present study aims to investigate the cytotoxic activity of Androctonus australis hector (Aah) scorpion venom and its toxic fractions (FtoxG-50 and F3) on NCI-H358 human lung cancer cells. Methods: The cytotoxic and antiproliferative activities were estimated using MTT assay, lactate dehydrogenase release and clonogenic assays. Apoptosis was evaluated by Hoechst 33258 staining, DNA fragmentation assay and caspase-3 activity. Oxidative stress was analyzed by reactive oxygen species, nitric oxide, malondialdehyde and protein carbonyl levels along with assessment of antioxidant status. In addition, alteration of mitochondrial membrane potential was analyzed by JC1 fluorescent dye. Results: The present findings showed that F3 fraction was more cytotoxic towards NCI-H358 lung cancer cells with an IC50 of 27.05 ± 0.70 μg/mL than venom alone (396.60 ± 1.33 μg/mL) and its toxic fraction FtoxG-50 (45.86 ± 0.91 μg/mL). Nevertheless, F3 fraction was not cytotoxic at these concentrations on normal human lung fibroblast MRC-5 cells. Inhibition of NCI-H358 cell proliferation after F3 fraction exposure occurred mainly by apoptosis as evidenced by damaged nuclei, significant DNA fragmentation level and caspase-3 activation in a dose dependent manner. Moreover, F3 fraction enhanced oxidative and nitrosative stress biomarkers and dissipated mitochondrial membrane potential in lung cancer cells along with significant depletion in cellular enzymatic and non-enzymatic antioxidants. Further, the apoptosis induced by F3 fraction was markedly prevented by the antioxidant N-acetylcysteine (NAC) suggesting the potential mechanism of oxidative stress. Conclusion: These findings suggest that F3 fraction could induce apoptosis in lung cancer cells through involvement of oxidative stress and mitochondrial dysfunction. Hence, these properties make F3 fraction a promising candidate for development of new anticancer agents.(AU)

Synthesis and activity evaluation of allyl-quercetin derivatives / 中国药学杂志

OBJECTIVE: To design and synthesize two kinds of allyl-substituted quercetin and investigate their anti-oxidation and anti-tumor activities in vitro. METHODS: The target compounds 1 and 2 were first synthesized from quercetin by hydroxyl protection, allylation,Claisen rearrangement and deprotection. The anti-oxidation and anti-tumor activities of the target compounds and intermediate products were evaluated by DPPH and MTT assay. RESULTS: Eight compounds were synthesized,including six intermediates and two target compounds,in which four were new compounds. All of them were confirmed by 1H-NMR,13C-NMR and LC-MS spectra. The anti-oxidation and anti-tumor tests showed that compounds 1,2,5,6,8 and 9 had anti-oxidation activities and compound 5 inhibited A549 lung cancer cell proliferation. Compound 9 could inhibit the proliferation of lung cancer A549 cells and HepG2 cells. CONCLUSION: The compounds with electron-donating groups have significant anti-oxidant activity. When acetyl and methyl ether groups are used as the protecting groups,the position of introducing allyl group to quercetin has obvious impact on the anti-tumor activity.

The research of mesoporous silica nanoparticles MCM-41 loading ginsenoside Rg3 for human lung cancer cells A549 / 中国药学杂志

OBJECTIVE: To improve the water solubility of ginsenoside Rg3, using mesoporous silica nanoparticles MCM-41 loading ginsenoside Rg3 and study the mechanism of promoting drug absorption with human lung cancer cells A549 as a model. METHODS: The mesoporous silica nanoparticles MCM-41, as a carrier, loading ginsenoside Rg3 by adsorption method. The morphology and particle size of MCM-41 were investigated by transmission electron microscopy (TEM) and laser particle size analyzer. The solid state characterization of ginsenoside Rg3 were investigated by differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD), and Fourier transform infrared spectroscopy method (FTIR), respectively. In vitro dissolution experiments investigated the dissolution rate of ginsenoside Rg3. Cytotoxicity assay and cell uptake experiments explored the effect of the administration system of A549 cells and the mechanism of inhibiting cell proliferation. RESULTS: We have been successfully prepared the mesoporous silica nanoparticles MCM-41. In vitro dissolution experiments showed that MCM-41 can significantly improve the dissolution rates of ginsenoside Rg3. Administration system can be uptaked into A549 cells and inhibit cell proliferation. CONCLUSION: The mesoporous silica nanoparticles MCM-41 has a good solubilization effect for ginsenoside Rg3, and MCM-41 has potential as a carrier for the treatment of lung cancer.