Journal: PloS one

Article Title: pp32 (ANP32A) expression inhibits pancreatic cancer cell growth and induces gemcitabine resistance by disrupting HuR binding to mRNAs.

doi: 10.1371/journal.pone.0015455

Figure Lengend Snippet: Figure 6. pp32 and HuR expression in clinical samples and patient outcomes. (A) The abundance and subcellular localization of HuR (left) and low to absent nuclear pp32 expression (right) in samples from pancreatic cancer patients were assessed by immunohistochemistry; magnification, 200x. Samples are representative of the cohort analyzed in B–D. (B) Correlation between pp32 nuclear expression and response to GEM treatment (p = 0.3, log rank test). (C) Correlation between high nuclear pp32-expressing tumor samples stratified into high or low HuR status in regards to GEM response (p = 0.88, log rank test). (D) Correlation between high cytoplasmic HuR-expressing tumor samples stratified into high and low pp32 nuclear expression correlated with GEM response (p = 0.25, log rank test). doi:10.1371/journal.pone.0015455.g006

Article Snippet: MiaPaCa2 pp32-transfected (Mia.pp32) and empty vector (Mia.EV) cells were trypsinized and collected as previously described [25] and our generated do novo using the previously generated and purchased parental pancreatic cancer cell line (ATCC, Manassas, VA).

Techniques: Expressing, Immunohistochemistry

Journal: Cancer Science

Article Title: Cancer‐testis antigen BORIS is a novel prognostic marker for patients with esophageal cancer

doi: 10.1111/j.1349-7006.2012.02355.x

Figure Lengend Snippet: Expression of BORIS m RNA in various cancer cell lines and cancer tissues and presence of BORIS ‐specific IgG in sera from patients with various cancers

Article Snippet: The cell lines used in the study were esophageal squamous cell carcinoma cell lines, TE2, TE3, TE4, TE5, TE6, TE7, TE8, TE9, TE10, TE11, TE12, TE13, TE14, and TE15 (Tohoku University, Sendai, Japan); melanoma cell lines, SKmel23, SKmel28, 888mel, A375mel, 1363mel, 928mel, 624mel, 501Amel, 586mel, 526mel, 501mel, 397mel, and 1362mel (Surgery Branch, NCI, NIH, Bethesda, MD, USA); colon cancer cell line, COLO205 (JCRB, Osaka, Japan); breast cancer cell line HS578 (American Type Culture Collection (ATCC), Manassas, VA, USA); stomach cancer cell lines, MKN1, MKN7, MKN28, MKN46, and MKN74 (Yamagata University, Yamagata, Japan); endometrial cancer cell line SNGII (Keio University, Tokyo, Japan); prostate cancer cell line LNCaP (ATCC); bladder cancer cell line, KU7 (Keio University); and brain tumor cell line U87MG (ATCC).

Techniques: Expressing

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 1 | Fra-1 inhibited the proliferation and promoted apoptosis of cervical cancer cells. (A) Cell viability of HeLa cells transfected with control vector (HeLa/vector) and HeLa cells transfected with Fra-1 overexpression vector (HeLa/Fra-1) cells determined by CCK8 assay. (B) Colony forming ability of HeLa/vector and HeLa/Fra-1 cells. (C) Colony numbers formed by HeLa/vector and HeLa/Fra-1 cells. (D) Cell cycle distribution among HeLa/vector and HeLa/Fra-1 cells as determined by flow cytometry. (E) Flow cytometric analysis of apoptosis among HeLa/vector and HeLa/Fra-1 cells. Data means ± standard deviation (SD). Each representative experiment was repeated three times with similar results. ∗∗P < 0.01; ∗∗∗P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Transfection, Control, Plasmid Preparation, Over Expression, CCK-8 Assay, Cytometry, Standard Deviation

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 2 | Fra-1 regulated p53 signaling pathway activity in cervical cancer cells. (A) mRNA chip analysis in HeLa/vector and HeLa/Fra-1 cells revealed differences in mRNA expression between the two groups. Differentially active signaling pathways were identified by gene ontology analysis. (B) Western blotting analysis of signal transducer and activator of transcription 1 (STAT1), p53, Bcl-2, p38, p21, MDM2, CDK4, and cyclinD1 protein levels in HeLa/vector and HeLa/Fra-1 cells. (C) Real-time polymerase chain reaction (PCR) analysis the mRNA expression levels of STAT1, p53, Bcl-2, p38, p21, MDM2, CDK4, and cyclinD1 in HeLa/vector and HeLa/Fra-1 cells. *P < 0.05; **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Activity Assay, Plasmid Preparation, Expressing, Protein-Protein interactions, Western Blot, Real-time Polymerase Chain Reaction

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 3 | Fra-1 promoted cell senescence and restored mitochondrial disorder in cervical cancer cells. (A) β-Galactosidase assay for cell senescence in HeLa/vector and HeLa/Fra-1 cells. (B) Western blot analysis of Fra-1, SIRT1, NF-κB, p16, and c-Myc protein expression in HeLa/vector and HeLa/Fra-1 cells. (C) Flow cytometric analysis of intracellular Ca2+ concentration. Red color represents HeLa/vector group and green color represents HeLa/Fra-1 group. (D) Flow cytometric analysis of intracellular reactive oxygen species (ROS) concentration. Red color represents HeLa/vector group and green color represents HeLa/Fra-1 group. (E) Flow cytometric analysis of mitochondrial membrane potential (1ψm) in HeLa/vector and HeLa/Fra-1 cells. (F) NAD+/NADH ratio in HeLa/vector and HeLa/Fra-1 cells. (G) Western blot analysis of sirtuin 3 (SIRT3), SOD2, IDH2, LKB1, and p-AMPK expression in HeLa/vector and HeLa/Fra-1 cells. **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Plasmid Preparation, Western Blot, Expressing, Concentration Assay, Membrane

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 5 | Silencing of signal transducer and activator of transcription 1(STAT1) by small interfering RNA promoted proliferation of cervical cancer cells via activation of the p53 signal pathway. (A) Real-time polymerase chain reaction (PCR) analysis of the silencing effects of three siRNAs for STAT1 in HeLa/Fra-1 cells. (B) Cell viability of HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells according to CCK8 assay. (C) Colony forming ability of HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (D) In vivo analysis of subcutaneous implanted tumor growth to compare the tumorigenic abilities of HeLa/vector, HeLa/Fra-1, and HeLa/Fra-1/siSTAT1 cells in mice. (E) Western blot analysis of protein expression of STAT1, CDK4, cyclin A, cyclin D1, p53, Bcl-2, c-REL, p38, p21, and MDM2 in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (F) Real-time PCR analysis of mRNA expression of STAT1, CDK4, cyclin D1, p53, Bcl-2, p38, p21, and MDM2 in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. *P < 0.05; **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Small Interfering RNA, Activation Assay, Real-time Polymerase Chain Reaction, CCK-8 Assay, In Vivo, Plasmid Preparation, Western Blot, Expressing

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 6 | Silencing of signal transducer and activator of transcription 1 (STAT1) inhibited senescence and promoted mitochondrial dysfunction in cervical cancer cells. (A) β-Galactosidase assay for cell senescence in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (B) Western blot analysis of protein expression of c-Myc, SIRT1, NF-κB, and p16 in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (C) Flow cytometric analysis of intracellular Ca2+ concentration. Red color represents HeLa/Fra-1 group and green color represents HeLa/Fra-1/siSTAT1 group. (D) Flow cytometric analysis of intracellular reactive oxygen species (ROS) levels. Red color represents HeLa/Fra-1 group and green color represents HeLa/Fra-1/siSTAT1 group. (E) MFI data reflecting changes in the intracellular ROS concentration in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (F) Flow cytometric analysis of mitochondrial membrane potential (1ψm) in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. *P < 0.05; **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Western Blot, Expressing, Concentration Assay, Membrane

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 7 | Silencing of signal transducer and activator of transcription 1(STAT1) recovered metabolic reprogramming in cervical cancer cells overexpressing Fra-1. (A) NAD + /NADH ratio in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (B) Western blot analysis of STAT1, sirtuin 3 (SIRT3), LKB1, and p-AMPK expression in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (C) Western blot analysis of the expression of key glycolysis enzymes PFK1, PKM2, and PDH in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (D) Real-time polymerase chain reaction (PCR) analysis of mRNA expression of glycolysis enzymes Glut1, HK II, and LDHA in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (E) Glucose concentration in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (F) Lactate concentration in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (G) Protein expression of glutamine dehydrogenase (GDH) and glutamine concentration in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. NS: No statistical difference; P < 0.05; **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Concentration Assay

Journal: The Journal of Clinical Investigation

Article Title: SIRT2 protects peripheral neurons from cisplatin-induced injury by enhancing nucleotide excision repair

doi: 10.1172/JCI123159

Figure Lengend Snippet: (A) Sirt2 KO in primary DRG neurons resulted in decreased cell survival after cisplatin treatment. Data were analyzed by 2-tailed Student’s t test (n = 3). (B and C) The viability of neuronally differentiated 50B11 (B) and PC12 (C) cells after various doses of cisplatin treatment was measured with trypan blue staining. Reexpression of WT-SIRT2, but not the enzymatically inactive mutant, HY-SIRT2, in Sirt2-KO 50B11 and PC12 cells increased cell resistance to cisplatin cytotoxicity. One-way ANOVA demonstrated a main effect for cell genotype (P < 0.001). *P < 0.05; **P < 0.01; ***P < 0.001 denote significance levels detected among various cell genotypes and at different time points by Tukey’s post hoc analysis. (D and E) The viability of control vector and Sirt2-KO LLC (D) and H1299 (E) cells after varying doses of cisplatin treatment, as measured by trypan blue staining. Sirt2 KO in LLC and H1299 cells shows no difference in cell viability following cisplatin treatment, as analyzed by 2-tailed Student’s t test.

Article Snippet: The mouse LLC cell line LL/2 (LLC1), human non–small cell lung cancer cell line H1299, and rat pheochromocytoma cell line PC12 were purchased from ATCC.

Techniques: Staining, Mutagenesis, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Establishment of zebrafish NSCLC BM xenograft models. ( A ) Schematic diagram of imaging and collecting zebrafish brain. The zebrafish brain was imaged from vertical view (blue arrow) and excised (red dotted box) to extract RNA. ( B ) The schematic diagram showed zebrafish embryos at 2 dpf. The red dots indicated cancer cells injected into the perivitelline space (PVs). ( C ) Images of brain at different developmental stages of Tg ( fli-1 : EGFP) zebrafish (2-6 dpf). The white short dashed line indicated the midbrain of the zebrafish, and the white long dashed line indicated the zebrafish’s eyes and hindbrain. ( D ) The expression of claudin-5 in the zebrafish brain at different developmental stages (2-7 dpf). ( E ) The expression of mfsd2aa and mfsd2ab in the zebrafish brain at different developmental stages (2-6 dpf). ( F ) Survival curves of 2 dpf zebrafish with different tumor-bearing amount in PVs. About 50-400 H1975 cells (red fluorescence) were injected into the PVs of 2 dpf zebrafish, and the number of deaths was counted till 8 dpi. ( G - I ) Three human NSCLC cell lines were involved: H1975, A549 and H1299. About 100 cells were injected into the PVs of zebrafish at different developmental stages, and the brain of zebrafish was imaged at 1 dpi. The white arrows indicated cancer cells in blood vessels of zebrafish brain. ( J - K ) Quantification of the BM cells number in zebrafish at 1 dpi. Significance was considered when P values were lower than 0.05. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001. dpf: days post fertilization, dpi: days post injection

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: Imaging, Injection, Expressing, Fluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: NSCLC cells could not proliferate in the midbrain of zebrafish at 4 dpi. ( A ) The schematic diagram showed zebrafish embryos at 2 dpf. The red dots indicated cancer cells injected into the midbrain of zebrafish. ( B ) Survival curves of zebrafish with different tumor-bearing amount at midbrain. About 50-400 H1975 cells (red fluorescence) were injected into the midbrain of 2 dpf Tg ( fli-1 : EGFP) zebrafish, and the number of deaths was counted till 8 dpi. ( C - G ) Five human cancer cell lines were involved: human breast cancer cell lines MDA-MB-231 (positive control) and MCF-7 (negative control), human NSCLC cancer lines H1975, A549 and H1299. About 100 cells were injected into the midbrain of zebrafish at 2 dpf, and the midbrain of zebrafish was imaged at 1 dpi and 4 dpi. (H-M) Quantification of cell proliferation in the midbrain of zebrafish at 1 dpi and 4 dpi. The fold change of cells in the midbrain was determined by dividing the measured values at 1 dpi and 4 dpi by the average measured values at 1 dpi. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: Injection, Fluorescence, Positive Control, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Zebrafish NSCLC BM xenograft models discriminated the BM potentials of different cell lines. ( A ) Transwell culture system was used to assess cell invasion ability in vitro , and five cancer cell lines were involved: MDA-MB-231, MCF-7, H1975, A549 and H1299. Invaded cells were stained with crystal violet (0.5 %) and imaged after 24 h incubation. ( B ) Quantification and comparison of invaded cells in vitro . Colonies were quantified using Image Pro Plus. ( C ) Five cancer cell lines were involved: MDA-MB-231, MCF-7, H1975, A549 and H1299. About 100 cells (red fluorescence) were injected into the PVs of Tg ( fli-1 : EGFP) zebrafish at 2 dpf, and the brain of zebrafish was imaged at 4 dpi. The white arrows indicated cancer cells in blood vessels of zebrafish brain. ( D ) Quantification of BM rate at 4 dpi. The number of BM cells in the same zebrafish at 4 dpi was divided by the number of BM cells at 1 dpi. The ratio of these two was named as BM potential. If the BM potential was greater than 1, it was considered that the zebrafish had brain metastasis. ( E - I ) Quantification of BM cells at 1 dpi and 4 dpi. The two dots connected by a straight line represented the number of BM cells of the same zebrafish at 1 dpi and 4 dpi. ( J ) Quantification and comparison of cell BM potential. The BM potential was determined by dividing the number of BM cells in the same zebrafish at 4 dpi by the number of BM cells at 1 dpi. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: In Vitro, Staining, Incubation, Comparison, Fluorescence, Injection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Zebrafish NSCLC BM xenograft models simultaneously discriminated the BM potentials of different cell lines. ( A ) The schematic diagram showed zebrafish embryos at 2 dpf. The red dots and green dots indicated cancer cells labeled with two different dyes were co-injected into the PVs of zebrafish. ( B ) About 50 H1975 cells (red fluorescence) and 50 H1975 cells (green fluorescence) were co-injected into the PVs of wild zebrafish at 2 dpf, and the brain of zebrafish was imaged at 4 dpi. The white long dashed line indicated the brain of zebrafish. ( C ) About 50 H1975 cells (red fluorescence) and 50 A549 cells (green fluorescence) were co-injected into the PVs of wild zebrafish at 2 dpf, and the brain of zebrafish was imaged at 4 dpi. ( D - G ) Quantification of BM cells in co-injection at 1 dpi and 4 dpi. The two dots connected by a straight line represented the number of BM cells of the same zebrafish at 1 dpi and 4 dpi. ( H and I ) The percentage of cells with two different dyes in the brain of zebrafish. The percentage of the cell line (red/green fluorescence) at 1 dpi was determined by dividing the number of BM cells (red/green fluorescence) by the total number of BM cells at 1 dpi. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: Labeling, Injection, Fluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Expression of microRNA-330-3p (miR-330-3p) affected the NSCLC BM potential in zebrafish xenograft models. ( A and B ) Quantification of the miR-330-3p expression in vitro . The appropriate transfection concentration was determined between 1 nM and 50 nM, according to the instructions. ( C and D ) The expression of miR-330-3p in H1975 cell line was knocked down by anti-miR-330-3p inhibitor and the expression of miR-330-3p in A549 cell line was overexpressed by over-miR-330-3p mimics. Transwell culture system was used to evaluate the effectiveness of transfection in vitro . Invaded cells were stained with crystal violet (0.5 %) and imaged after 24 h incubation. ( E and F ) Quantification of invaded cells in vitro . Colonies were quantified using Image Pro Plus. ( G and H ) H1975, H1975 with under-expressed miR-330-3p, A549 and A549 with over-expressed miR-330-3p were involved. About 100 cells (red fluorescence) were injected into the PVs of Tg ( fli-1 : EGFP) zebrafish at 2 dpf to evaluate the effectiveness of transfection in vivo , and the brain of zebrafish was imaged at 4 dpi. The white arrows indicated cancer cells in blood vessels of zebrafish brain. ( I - L ) Quantification of BM cells at 1 dpi and 4 dpi. The two dots connected by a straight line represented the number of BM cells of the same zebrafish at 1 dpi and 4 dpi. ( M and N ) Quantification and comparison of the BM potentials of the transfected cell lines and the original cell lines. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: Expressing, In Vitro, Transfection, Concentration Assay, Staining, Incubation, Fluorescence, Injection, In Vivo, Comparison

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Zebrafish NSCLC BM xenograft models discriminated different chemosensitivities within 4 days. ( A and B ) NSCLC cell lines H1975 and A549 were involved. Transwell culture system was used to evaluate the effectiveness of osimertinib (1 µM) and gefitinib (2 µM) against cell invasion in vitro . Invaded cells were stained with crystal violet (0.5 %) and imaged after 24 h incubation. ( C and D ) Quantification of invaded cells in vitro. Colonies were quantified using Image Pro Plus. ( E and F ) NSCLC cell lines H1975 and A549 were involved. Tg ( fli-1 : EGFP) zebrafish embryos were injected about 100 cells (red fluorescence) into the PVs at 2 dpf and administered with osimertinib (1 µM) and gefitinib (13 µM) by intracardiac injection. ( G and H ) Quantification of cell brain metastasis at 4 dpi. Fold change of BM cell number was determined by dividing the number of BM cells in the same zebrafish at 4 dpi by the number of BM cells at 1 dpi. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: In Vitro, Staining, Incubation, Injection, Fluorescence