Recombinant adeno-associated virus 9-mediated expression of Kallistatin suppresses lung tumor growth in mice

Background:: Lung cancer remains the most common cause of cancer-related deaths in China and worldwide. Traditional surgery and chemotherapy do not offer an effective cure although gene therapy may be a promising future alter-native. Kallistatin (Kal) is an endogenous inhibitor of angiogenesis and tumorigenesis. Recombinant adeno-associated virus (rAAV) is considered the most promising vector for gene therapy of many diseases due to persistent and long-term transgen-ic expression. Objective:: The aim of this study was to investigate whether rAAV9-Kal inhibited NCI-H446 subcutaneous xenograft tumor growth in mice. Method:: The subcutaneous xenograft mode were induced by subcutaneous injection of 2×106 H446 cells into the dorsal skin of BALB/c nude mice. The mice were administered with ssrAAV9-Kal (single-stranded rAAV9) or dsrAAV9-Kal (double-stranded rAAV9）by intraperitoneal injection (I.P.). Tumor microvessel density (MVD) was examined by anti-CD34 stain-ing to evaluate tumor angiogenesis. Results:: Compared with the PBS (blank control) group, tumor growth in the high-dose ssrAAV9-Kal group was inhibited by 40% by day 49, and the MVD of tumor tissues was significantly decreased. Conclusion:: The results indicate that this therapeutic strategy is a promising approach for clinical cancer therapy and impli-cate rAAV9-Kal as a candidate for gene therapy of lung cancer.

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Antitumor efficacy of 2-(6,8-dimethyl-5-nitro-4-chloroquinoline-2-yl)-5,6,7-trichloro-1,3-tropolone in lung cancer xenografts.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.e15008 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. e15008-e15008

Author(s):

Ekaterina A. Lukbanova ◽

Anna S. Goncharova ◽

A. V. Volkova ◽

M. V. Mindar ◽

Ekaterina V. Zaikina ◽

...

Keyword(s):

Lung Cancer ◽

Tumor Growth ◽

A549 Cells ◽

Cytotoxic Agents ◽

Antitumor Efficacy ◽

Control Group ◽

Common Cancer ◽

Subcutaneous Xenograft ◽

Starch Gel ◽

Inhibition Of Tumor Growth

e15008 Background: Lung cancer is the most common cancer worldwide. The efficacy of chemotherapy for this tumor does not exceed 40%. Moreover, all cytotoxic agents cause many side effects. The search for new substances with an antitumor effect seems to be relevant. Tropolone alkaloids, which are seven-membered non-benzenoid aromatic compounds, are promising inhibitors of tumor growth. The purpose of the study was to evaluate the antitumor efficacy of 2-(6,8-dimethyl-5-nitro-4-chloroquinoline-2-yl)-5,6,7-trichloro-1,3-tropolone in subcutaneous xenografts of lung cancer A549 cells in immunodeficient Balb/c Nude mice. Methods: The study was performed on a PDX model of lung cancer created by subcutaneous injection of A549 cell suspension, 5x106 cells in 0.2 ml of a solution of serum-free nutrient medium 199 and Matrigel (1:1). Animals were equally divided into 5 groups (each n = 5). Experimental groups 1, 2, 3 and 4 received 2-(6,8-dimethyl-5-nitro-4-chloroquinoline-2-yl)-5,6,7-trichloro-1,3-tropolone at doses of 0.0055, 0.055, 0.55 and 2.75 mg/g, respectively; the control group received 1% starch gel. The dynamics of the growth of a subcutaneous xenograft was evaluated by measuring tumor nodes. The volumes of tumor nodes were calculated using the Shrek’s formula for the ellipsoid: V = a×b×c×p/6, where V is the tumor volume (mm3), and a, b, c are the maximum dimensions of the ellipsoid in three planes (mm). The data were statistically analyzed using the Exel and Medstatistic progams. Results: The dynamics of subcutaneous xenograft growth in groups 1, 2, 3 and 4 significantly differed from the control group (p = 0.023). The average tumor volumes on the 39th day after the implantation in the control group and in experimental groups 1, 2, 3 and 4 were 1159.2, 895.3, 565.8, 80.7 and 76.7 mm3, respectively. The inhibition of tumor growth (ITG) was in direct proportion to the dose of the administered substance. Lower ITG(%) (32.4, 23.2 and 10.3% in experimental groups 2, 3 and 4, respectively) was associated with lower concentrations of О154 (0.55 mg/g, 0.055 mg/g and 0.0055 mg/g, respectively). Conclusions: The study demonstrated statistically significant differences in xenograft volume indices in all experimental groups compared to the control group.

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Assessment of an antitumor effect of 2-(6,8-dimethyl-5-nitro-4-chloroquinoline2-yl)-5,6,7-trichloro-1,3-tropolone in A-549 tumor cell subcutaneous xenografts

Almanac of Clinical Medicine ◽

10.18786/2072-0505-2021-49-021 ◽

2020 ◽

Vol 49 ◽

Author(s):

E. A. Lukbanova ◽

E. V. Zaikina ◽

Yu. A. Sayapin ◽

E. A. Gusakov ◽

S. Yu. Filippova ◽

...

Keyword(s):

Lung Cancer ◽

Tumor Growth ◽

Tumor Cells ◽

Nude Mice ◽

Antitumor Effect ◽

Lung Tumor ◽

Cytotoxic Agents ◽

Effective Dosage ◽

Control Group ◽

Group 5

Rationale: Chemotherapy is one of the lung cancer treatment methods. The search for new substances with antitumor effect against malignant lung neoplasms is relevant because of low efficacy and side effects of cytotoxic agents. A promising substance class with various biological activities, including antitumor, includes alkaloids of the tropolone family, such as heptamerous non-benzoid aromatic compounds. 2-(6,8-dimethyl-5-nitro-4-chloroquinoline-2-yl)- 5,6,7-trichloro-1,3-tropolone has been synthesized in Institute of Physical and Organic Chemistry; it is a new compound belonging to 2-quinoline-2-yl derivatives of 1,3-tropolone.Aim: To assess the antitumor effect of 2-(6,8-dimethyl-5-nitro-4-chloroquinoline-2-yl)- 5,6,7-trichloro-1,3-tropolone on subcutaneous xenografts of A-549 lung tumor cells in immunodeficient Balb/c Nude mice.Materials and methods: The study included 50 immunodeficient Balb/c Nude mice divided into 4 experimental groups depending on the dosage of the study substance (0.0055, 0.055, 0.55, and 2.75 mg/g); group 5 was the control group. A-549 cells of lung cancer were used as a xenograft. The antitumor effect of tropolone was evaluated by the inhibition of tumor growth and the index of tumor growth. The experiment lasted for 36 days starting from the first administration of the substances.Results: The mean tumor volumes on day 36 of the experiment in the control group and four experimental groups were 2729.5; 2150.8; 1746.4; 952.3 and 678.9 mm3 , respectively. The indices of tumor growth in groups 1, 2, 3 and 4 were significantly lower than in group 5 (control) starting from days 24, 21, 21 and 15, respectively, and till the end of the experiment. Maximal differences between groups 4 and 5 were observed at days 33 and 36 (by 3.7, p=0.01 and 4.1, p=0.003 times, respectively).Discussion: The anti-tumor effect of 2-(6,8-dimethyl-5-nitro-4-chloroquinoline-2-yl)- 5,6,7-trichloro-1,3-tropolone demonstrated in the study could be related to various mechanisms. For example, numerous studies have shown that its related compound hinokitiol exerts a cytotoxic effect associated with cessation of the cell cycle, apoptosis induction, DNA damage, and autophagic death of tumor cells.Conclusion: The study demonstrated significant differences in xenograft volumes in all experimental groups, compared to the control group. In mice, 2.75 mg/g bodyweight was the most effective dosage of the studied compound leading to a slow decrease in tumor growth rates and a decrease in the volumes of subcutaneous xenografts.

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Kras-driven intratumoral heterogeneity triggers infiltration of M2 polarized macrophages via the circHIPK3/PTK2 immunosuppressive circuit

Scientific Reports ◽

10.1038/s41598-021-94671-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Theodora Katopodi ◽

Savvas Petanidis ◽

Kalliopi Domvri ◽

Paul Zarogoulidis ◽

Doxakis Anestakis ◽

...

Keyword(s):

Lung Cancer ◽

Tumor Growth ◽

Lung Tumor ◽

Quantitative Polymerase Chain Reaction ◽

Macrophage Polarization ◽

Metastatic Potential ◽

Intratumoral Heterogeneity ◽

M2 Macrophage

AbstractIntratumoral heterogeneity in lung cancer is essential for evasion of immune surveillance by tumor cells and establishment of immunosuppression. Gathering data reveal that circular RNAs (circRNAs), play a role in the pathogenesis and progression of lung cancer. Particularly Kras-driven circRNA signaling triggers infiltration of myeloid-associated tumor macrophages in lung tumor microenvironment thus establishing immune deregulation, and immunosuppression but the exact pathogenic mechanism is still unknown. In this study, we investigate the role of oncogenic Kras signaling in circRNA-related immunosuppression and its involvement in tumoral chemoresistance. The expression pattern of circRNAs HIPK3 and PTK2 was determined using quantitative polymerase chain reaction (qPCR) in lung cancer patient samples and cell lines. Apoptosis was analyzed by Annexin V/PI staining and FACS detection. M2 macrophage polarization and MDSC subset analysis (Gr1−/CD11b−, Gr1−/CD11b+) were determined by flow cytometry. Tumor growth and metastatic potential were determined in vivo in C57BL/6 mice. Findings reveal intra-epithelial CD163+/CD206+ M2 macrophages to drive Kras immunosuppressive chemoresistance through myeloid differentiation. In particular, monocytic MDSC subsets Gr1−/CD11b−, Gr1−/CD11b+ triggered an M2-dependent immune response, creating an immunosuppressive tumor-promoting network via circHIPK3/PTK2 enrichment. Specifically, upregulation of exosomal cicHIPK3/PTK2 expression prompted Kras-driven intratumoral heterogeneity and guided lymph node metastasis in C57BL/6 mice. Consequent co-inhibition of circPTK2/M2 macrophage signaling suppressed lung tumor growth along with metastatic potential and prolonged survival in vivo. Taken together, these results demonstrate the key role of myeloid-associated macrophages in sustaining lung immunosuppressive neoplasia through circRNA regulation and represent a potential therapeutic target for clinical intervention in metastatic lung cancer.

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Systemic Platelet-activating Factor Receptor Activation Augments Experimental Lung Tumor Growth and Metastasis

Cancer Growth and Metastasis ◽

10.4137/cgm.s14501 ◽

2014 ◽

Vol 7 ◽

pp. CGM.S14501 ◽

Cited By ~ 15

Author(s):

Patrick C. Hackler ◽

Sarah Reuss ◽

Raymond L. Konger ◽

Jeffrey B. Travers ◽

Ravi P. Sahu

Keyword(s):

Lung Cancer ◽

Tumor Growth ◽

Cancer Progression ◽

Lung Tumor ◽

Platelet Activating Factor ◽

Receptor Activation ◽

Dependent Manner ◽

Melanoma Tumor ◽

Tumor Growth And Metastasis ◽

The Impact

Pro-oxidative stressors including cigarette smoke (CS) generate novel lipids with platelet-activated factor-receptor (PAF-R) agonistic activity mediate systemic immunosuppression, one of the most recognized events in promoting carcinogenesis. Our previous studies have established that these oxidized-PAF-R-agonists augment murine B16F10 melanoma tumor growth in a PAF-R-dependent manner because of its effects on host immunity. As CS generates PAF-R agonists, the current studies sought to determine the impact of PAF-R agonists on lung cancer growth and metastasis. Using the murine Lewis Lung Carcinoma (LLC1) model, we demonstrate that treatment of C57BL/6 mice with a PAF-R agonist augments tumor growth and lung metastasis in a PAF-R-dependent manner as these findings were not seen in PAF-R-deficient mice. Importantly, this effect was because of host rather than tumor cells PAF-R dependent as LLC1 cells do not express functional PAF-R. These findings indicate that experimental lung cancer progression can be modulated by the PAF system.

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Deep Learning Based Automated Orthotopic Lung Tumor Segmentation in Whole-Body Mouse CT-Scans

Cancers ◽

10.3390/cancers13184585 ◽

2021 ◽

Vol 13 (18) ◽

pp. 4585

Author(s):

Wouter R. P. H. van de Worp ◽

Brent van der Heyden ◽

Georgios Lappas ◽

Ardy van Helvoort ◽

Jan Theys ◽

...

Keyword(s):

Lung Cancer ◽

Deep Learning ◽

Tumor Growth ◽

Tumor Volume ◽

Learning Algorithm ◽

Lung Tumor ◽

Whole Body ◽

Ct Scans ◽

Therapeutic Effects ◽

Deep Learning Algorithm

Lung cancer is the leading cause of cancer related deaths worldwide. The development of orthotopic mouse models of lung cancer, which recapitulates the disease more realistically compared to the widely used subcutaneous tumor models, is expected to critically aid the development of novel therapies to battle lung cancer or related comorbidities such as cachexia. However, follow-up of tumor take, tumor growth and detection of therapeutic effects is difficult, time consuming and requires a vast number of animals in orthotopic models. Here, we describe a solution for the fully automatic segmentation and quantification of orthotopic lung tumor volume and mass in whole-body mouse computed tomography (CT) scans. The goal is to drastically enhance the efficiency of the research process by replacing time-consuming manual procedures with fast, automated ones. A deep learning algorithm was trained on 60 unique manually delineated lung tumors and evaluated by four-fold cross validation. Quantitative performance metrics demonstrated high accuracy and robustness of the deep learning algorithm for automated tumor volume analyses (mean dice similarity coefficient of 0.80), and superior processing time (69 times faster) compared to manual segmentation. Moreover, manual delineations of the tumor volume by three independent annotators was sensitive to bias in human interpretation while the algorithm was less vulnerable to bias. In addition, we showed that besides longitudinal quantification of tumor development, the deep learning algorithm can also be used in parallel with the previously published method for muscle mass quantification and to optimize the experimental design reducing the number of animals needed in preclinical studies. In conclusion, we implemented a method for fast and highly accurate tumor quantification with minimal operator involvement in data analysis. This deep learning algorithm provides a helpful tool for the noninvasive detection and analysis of tumor take, tumor growth and therapeutic effects in mouse orthotopic lung cancer models.

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Spice-derived phenolic, malabaricone B induces mitochondrial damage in lung cancer cellsviaa p53-independent pathway

Food & Function ◽

10.1039/c8fo00624e ◽

2018 ◽

Vol 9 (11) ◽

pp. 5715-5727 ◽

Cited By ~ 1

Author(s):

Mrityunjay Tyagi ◽

Biswanath Maity ◽

Bhaskar Saha ◽

Ajay Kumar Bauri ◽

Mahesh Subramanian ◽

...

Keyword(s):

Lung Cancer ◽

Cell Death ◽

Tumor Growth ◽

Lung Tumor ◽

Mitochondrial Damage

The spice-derived phenolic, malabaricone B induces mitochondrial cell death and reduces lung tumor growthin vivo.

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Gingerol inhibits cisplatin-induced acute and delayed emesis in rats and minks by regulating the central and peripheral 5-HT, SP, and DA systems

Journal of Natural Medicines ◽

10.1007/s11418-019-01372-x ◽

2019 ◽

Vol 74 (2) ◽

pp. 353-370 ◽

Cited By ~ 2

Author(s):

Li Tian ◽

Weibin Qian ◽

Qiuhai Qian ◽

Wei Zhang ◽

Xinrui Cai

Keyword(s):

Low Dose ◽

Area Postrema ◽

Biologically Active ◽

Control Group ◽

High Dose ◽

Delayed Emesis ◽

Blank Control Group ◽

Qrt Pcr ◽

Underlying Mechanisms ◽

Different Doses

Abstract Gingerol, a biologically active component in ginger, has shown antiemetic properties. Our study aimed to explore the underlying mechanisms of gingerol on protecting rats and minks from chemotherapy-induced nausea and vomiting. The preventive impact of gingerol was evaluated in the pica model of rats and the vomiting model of minks induced by cisplatin at every 6 h continuously for a duration of 72 h. Animals were arbitrarily separated into blank control group, simple gingerol control group, cisplatin control group, cisplatin + metoclopramide group, cisplatin + three different doses gingerol group (low-dose; middle-dose; high-dose). The area postrema as well as ileum damage were assessed using H&E stain. The levels of 5-TH, 5-HT3 receptor, TPH, SERT, SP, NK1 receptor, PPT, NEP, DA, D2R, TH, and DAT were determined using immunohistochemistry or qRT-PCR in rats and minks. All indicators were measured in the area postrema along with ileum. The kaolin intake by rats and the incidence of CINV of minks were significantly decreased after pretreatment with gingerol in a dosage-dependent way for the duration of 0–24-h and 24–72-h. Gingerol markedly decreased the levels of 5-TH, 5-HT3 receptor, TPH, SP, NK1 receptor, PPT, DA, D2R, TH, alleviated area postrema as well as ileum damage, and increased the accumulation of SERT, NEP, DAT in the area postrema along with ileum of rats and minks. Gingerol alleviates cisplatin-induced kaolin intake of rats and emesis of minks possibly by regulating central and peripheral 5-HT system, SP system and DA system. Graphic abstract

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Tumor-Derived CXCL1 Promotes Lung Cancer Growth via Recruitment of Tumor-Associated Neutrophils

Journal of Immunology Research ◽

10.1155/2016/6530410 ◽

2016 ◽

Vol 2016 ◽

pp. 1-11 ◽

Cited By ~ 19

Author(s):

Ming Yuan ◽

Ha Zhu ◽

Junfang Xu ◽

Yuanyuan Zheng ◽

Xuetao Cao ◽

...

Keyword(s):

Lung Cancer ◽

Tumor Growth ◽

Cancer Biology ◽

First Line ◽

Traditional Role ◽

Tumor Tissues ◽

Tumorigenesis And Progression ◽

Tumor Associated Neutrophils ◽

Cxcl1 Expression

Neutrophils have a traditional role in inflammatory process and act as the first line of defense against infections. Although their contribution to tumorigenesis and progression is still controversial, accumulating evidence recently has demonstrated that tumor-associated neutrophils (TANs) play a key role in multiple aspects of cancer biology. Here, we detected that chemokine CXCL1 was dramatically elevated in serum from 3LL tumor-bearing mice. In vitro, 3LL cells constitutively expressed and secreted higher level of CXCL1. Furthermore, knocking down CXCL1 expression in 3LL cells significantly hindered tumor growth by inhibiting recruitment of neutrophils from peripheral blood into tumor tissues. Additionally, tumor-infiltrated neutrophils expressed higher levels of MPO and Fas/FasL, which may be involved in TAN-mediated inhibition of CD4+ and CD8+ T cells. These results demonstrate that tumor-derived CXCL1 contributes to TANs infiltration in lung cancer which promotes tumor growth.

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Inhibitory effect of endostar combined with radiotherapy on gastric cancer animal models

10.21203/rs.3.rs-19229/v2 ◽

2020 ◽

Author(s):

Qitian Chen ◽

Ran Chen ◽

Youhong Dong

Keyword(s):

Tumor Growth ◽

Inhibitory Effect ◽

The Body ◽

Combination Group ◽

Control Group ◽

Blank Control Group ◽

Expression Levels ◽

Radiotherapy Group ◽

Significant Difference ◽

Tgf Β1

Abstract Background: Inhibitory effect of endostar combined with radiotherapy on gastric cancer (GC) animal models and its effect on transforming growth factor-β1 (TGF-β1) and inter- leukin-10 (IL-10) were evaluated. Methods: Forty mice of a GC model xenograft tumors were prepared and randomly divided into blank control group, endostar group, radiotherapy group and endostar combined with radiotherapy group (combination group). From the 14th day, a vernier caliper was used for measuring the long and short diameters of the xenograft tumors. The formula V = ab2/2 was used for calculating the tumor volume and to obtain its average value. Tumor growth curves were plotted to calculate the tumor inhibition rate. The growth of xenograft tumors and the behavioral changes of mice were observed. Enzyme-linked immunosorbent assay (ELISA) was used for detecting the expression levels of IL-10 and TGF-β1. Results: The tumor growth in the combination group was significantly inhibited and the tumor volume was the smallest compared with the other groups (p<0.05). Compared to the blank control group, the tumor inhibition rate was 11.8% in endostar group, 33.0% in radiotherapy group and 52.1% in combination group (p<0.01). Endostar combined with radiotherapy had an interaction in decreasing the expression levels of TGF-β1 and IL‑10 (F=4.35 and 5.12, p<0.05). Leucocyte count was significantly higher in control and combination groups than that in endostar and radiotherapy groups. The body weight of mice in endostar and radiotherapy groups decreased after treatment (p<0.05). The body weight of mice after treatment in control and combination groups increased, with a statistically significant difference compared to that before treatment (p<0.05). There was a statistically significant difference among all groups after treatment (F=198.1, p<0.01). Conclusions: Endostar combined with radiotherapy can inhibit tumor growth and downregulate the expression levels of TGF-β1 and IL-10 through synergistic action.

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Dexmedetomidine Associated Lung Cancer Proliferation and Tumor Growth is Prevented by β-Caryophyllene: Role of AMPK Activation and PGC-1α/TFAM Over Expression

Indian Journal of Animal Research ◽

10.18805/ijar.bf-1431 ◽

2022 ◽

Author(s):

Dan Wang ◽

Dazhi Long ◽

Jiegang Zhou ◽

Ziqiang Dong ◽

Guiming Huang

Keyword(s):

Lung Cancer ◽

Cell Proliferation ◽

Tumor Growth ◽

Lung Tumor ◽

Apoptotic Cell ◽

Tumor Development ◽

A549 Cells ◽

Treated Group ◽

Metastatic Progression ◽

Cancer Proliferation

Background: Dexmedetomidine has been reported to induce anti-apoptotic effects and metastatic progression in lung cancer. In the current investigation, the effect of β-Caryophyllene on dexmedetomidine induced cell proliferation and apoptosis of lung cancer cells and tumor growth in mice was studied. Methods: A549 cell line was cultured with either dexmedetomidine alone or together with β-Caryophyllene for 24 h and analysed for cell proliferation with MTT assay. ELISA based kit was used to determine apoptotic DNA fragmentation. Western blotting was used to determine expression levels of target proteins. The induction of experimental lung tumor in rat model was achieved through the injection of A549 tumor cells subcutaneously into the middle left side of the mice after anesthetization with pentobarbital (35 mg/kg) at 2.8 × 106 cells in 400 μl of PBS. Result: We found that β-Caryophyllene exerts the anti-proliferative effects on A549 cells. Furthermore, β-Caryophyllene significantly prevents apoptotic cell death and causes up-regulation of PGC-1α and TFAM compared to dexmedetomidine treated cells. We observed that β-Caryophyllene suppressed tumor development in mice significantly compared to dexmedetomidine treated group without changing body weight.

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