Establishment of Peritoneal and Hepatic Metastasis Mouse Xenograft Models Using Gastric Cancer Cell Lines

Miwa, Takashi; Kanda, Mitsuro; Umeda, Shinichi; Tanaka, Haruyoshi; Shimizu, Dai; Tanaka, Chie; Kobayashi, Daisuke; Hayashi, Masamichi; Yamada, Suguru; Nakayama, Goro; Koike, Masahiko; Kodera, Yasuhiro

doi:10.21873/invivo.11669

Cited by 21 publications

(16 citation statements)

References 35 publications

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“…With regard to tumorigenic ability of KatoIII cells in nude mice, contradictory results have been reported. While several groups demonstrated the tumorigenic capacity of KatoIII cells in nude mice [ 43 – 46 ], others failed to develop the tumor in nude mice [ 47 , 48 ]. We do not know a precise reason for the inconsistent reports attributed to the KatoIII tumorigenicity in nude mice.…”

Section: Discussionmentioning

confidence: 99%

miR-34a mimic or pre-mir-34a, which is the better option for cancer therapy? KatoIII as a model to study miRNA action in human gastric cancer cells

2021

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Background Aberrantly expressed microRNAs play important roles in gastric tumorigenesis. However, use of miRNAs as a therapeutic option in gastric cancer still remains as a challenging problem. Methods We performed transient transfection of miR-34a-5p mimic and stable transfection of pre-mir-34a into KatoIII cells. Then, we evaluated the effect of transfected miRNAs on numerous cellular and molecular processes. Results Following transient transfection of miR-34a-5p mimic at 25 nM—a commonly used concentration—into KatoIII cells, inhibition of two target genes expression, namely Notch1 and β-catenin, was not observed, but a non-significant marginal increase of these genes was detected. No changes were detected in the percentage of apoptotic cells as well as in CD44 + and EpCAM + cells after 25 nM miR-34a-5p mimic transfection. Interestingly, stable transfection of pre-mir-34a into KatoIII cells (named as KatoIII-pGFPC1-34a cells) caused a significant repression in β-catenin protein and Notch1 mRNA levels (p < 0.05 and p < 0.01, respectively) relative to equivalent control (KatoIII- pGFPC1-empty cells). The percentage of CD44 + cells in the KatoIII-pGFPC1-34a cells (< 40%) was significantly lower than that in control cells (~ 95%) (p < 0.05). An increase of ~ 3.5% in apoptotic cells and a slower proliferation rate were detected in KatoIII-pGFPC1-34a cells. Conclusions Our study revealed that the effect of miR mimic in target gene repression can be dependent to its concentration as well as to the cell type. Meanwhile, our findings further support a regulatory function for pre-miRNAs in target repression and will help to develop effective therapeutic strategies in cancer treatment.

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Section: Discussionmentioning

confidence: 99%

miR-34a mimic or pre-mir-34a, which is the better option for cancer therapy? KatoIII as a model to study miRNA action in human gastric cancer cells

2021

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“…To modulate NPTXR expression, we generated GC cell lines with small interfering RNA (siRNA)-mediated KD, short hairpin RNA (shRNA)-mediated KD, CRISPR-Cas9-mediated stable KO, and forced overexpression (see Additional file 2: Table S1 1 for sequence details). Genome editing using the CRISPR-Cas9 system was used to generate stable NPTXR-KO GC cell lines from MKN1 cells stably expressing luciferase because it expressed one of the highest levels of NPTXR mRNA, had high abilities in cell migration and invasion in our previous studies and is engrafted in nude mice for subcutaneous and peritoneal xenograft models [15,16].…”

Section: Nptxr Knockdown (Kd) Knockout (Ko) and Overexpression In Gmentioning

confidence: 99%

Therapeutic monoclonal antibody targeting of neuronal pentraxin receptor to control metastasis in gastric cancer

et al. 2020

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Background: Controlling metastasis is essential for improving the prognosis of patients with gastric cancer (GC). Here, we aimed to identify a molecule required for GC metastasis and to investigate its potential utility as a target for the development of therapeutic antibodies (Abs). Methods: Transcriptome and bioinformatics analyses of human GC cell lines identified the neuronal pentraxin receptor (NPTXR) as a candidate molecule. NPTXR function was probed by modulating its expression in GC cells and assessing the effects on intracellular signaling and malignant behaviors in vitro and in mouse xenograft models. We also generated anti-NPTXR Abs and Nptxr −/− mice, and assessed the clinical significance of NPTXR expression in GC specimens. Results: NPTXR mRNA expression in clinical specimens was associated with disease progression and was significantly higher in tissues from GC patients with distant metastasis compared with those without. NPTXR regulated expression of genes involved in metastatic behaviors as well as activation of the PI3K-AKT-mTOR, FAK-JNK, and YAP signaling pathways. NPTXR silencing promoted caspase-mediated apoptosis and attenuated GC cell proliferation, cell cycle progression, migration, invasion, adhesion, stem cell-like properties, and resistance to 5fluorouracil in vitro, and also inhibited the tumorigenicity of GC cells in vivo. Anti-NPTXR Abs inhibited GC peritoneal metastasis in mice. Nptxr −/− mice showed no abnormalities in reproduction, development, metabolism, or motor function. Conclusions: NPTXR plays an essential role in controlling the malignant behavior of GC cells in vitro and in vivo. NPTXR-targeting Abs may thus have utility as novel diagnostic tools and/or treatment modalities for GC.

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“…Animal experiments were conducted using 6-week-old female Balb/c nude mice [ 18 ]. A total of 1 × 10 7 DNAJC19-knockdown A549 cells or negative control A549 cells were suspended in 100 µl of phosphate-buffered saline and subcutaneously injected into each mouse to establish the xenograft model; every group had 10 mice.…”

Section: Methodsmentioning

confidence: 99%

Targeting DNAJC19 overcomes tumor growth and lung metastasis in NSCLC by regulating PI3K/AKT signaling

Zhou

Yang

Gao³

et al. 2021

Cancer Cell Int

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Background Some driver oncogenes are still unknown in non-small-cell lung cancer (NSCLC). DNAJC19, a major component of the translocation machinery of mitochondrial membranes, is a disease-associated protein. Herein, we report the role of DNAJC19 in NSCLC cell growth and metastasis. Methods Immunohistochemistry (IHC) was performed to investigate DNAJC19 expression in NSCLC clinical samples. For knockdown or overexpression assays in A549 or NCI-H1299 lung cancer cells, lentiviral vectors were used. After assessment of cell functions, DNAJC19-knockdown A549 cells were further applied to establish mouse xenograft and metastasis tumor models. Assessments based on the RNA-seq data, western blotting, PCR and IHC were performed for the mechanistic study. Results Expression of DNAJC19 was higher in tumors than in noncancerous adjacent tissues. Survival analysis indicated that low DNAJC19 levels were correlated with an increased progression-free survival rate. ShRNA-mediated knockdown of DNAJC19 markedly inhibited cell growth, viability, migration and invasion. Moreover, RNA-seq analysis revealed that the PI3K/AKT signaling pathway was involved in molecular events when A549 cells were treated with shDNAJC19. In addition, DNAJC19 knockdown decreased PI3Kp85a, AKT and p-AKT expression in A549 cells, and cellular functions were greatly rescued in DNAJC19-knockdown A549 cells by ectopic overexpression of AKT. Furthermore, tumor xenograft growth and lung metastasis were markedly repressed in the shDNAJC19 group compared to the control group. As expected, the expression levels of DNAJC19, PI3K and AKT in xenograft mouse samples were also lower in the shDNAJC19 group than in the shCtrl group. Conclusions DNAJC19 greatly promotes NSCLC cell growth and lung metastasis by regulating PI3K/AKT signaling, providing a novel therapeutic target for treating NSCLC patients.

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Establishment of Peritoneal and Hepatic Metastasis Mouse Xenograft Models Using Gastric Cancer Cell Lines

Cited by 21 publications

References 35 publications

miR-34a mimic or pre-mir-34a, which is the better option for cancer therapy? KatoIII as a model to study miRNA action in human gastric cancer cells

miR-34a mimic or pre-mir-34a, which is the better option for cancer therapy? KatoIII as a model to study miRNA action in human gastric cancer cells

Therapeutic monoclonal antibody targeting of neuronal pentraxin receptor to control metastasis in gastric cancer

Targeting DNAJC19 overcomes tumor growth and lung metastasis in NSCLC by regulating PI3K/AKT signaling

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