Project description:Chinese lung adenocarcinoma cancer cells (SPC-A-1) and human larger cell lung cancer cells (NCI-H460) were injected into left cardiac ventricle of nude mice for bone metastases model, respectively. Bone metastatic lesions were detected by bone scintigraphy with 99mTc-methylene diphophonate, removed bone metastatic lesions for cell primary culture, chromosome analysis for determine the bone metastatic cells have a characterization of unchanged humanization, in the anesthesia death mice. Through eight in vivo ~ in vitro selections, the 4th, 8th generation cells of SPC-A-1, 8th generation cells of NCI-H460 and their parental cells were used for microarray analysis, respectively. Bone metastatic clones 4th and/or 8th generation SPC-A-1 vs. SPC-A-1, 8th generation NCI-H460 vs. NCI-H460, respectively. Biological replicates: one replicate for every group, independently grown and harvested. One replicate per array.
Project description:CRISPRi-mediated transcriptional inhibition of CPT1 with two distinct sgRNAs in NCI-H460 lung cancer cells, to investigate the dynamics of gene expression regulation upon CPT1 knockdown.
Project description:We conducted a proteomic analysis of secretome and EV samples isolated from the conditioned medium of lung cancer LC (A549, NCI-H23, and NCI-H460) and CRC (Caco2, HCT116, and HT-29) cell line models.
Project description:Chinese lung adenocarcinoma cancer cells (SPC-A-1) and human larger cell lung cancer cells (NCI-H460) were injected into left cardiac ventricle of nude mice for bone metastases model, respectively. Bone metastatic lesions were detected by bone scintigraphy with 99mTc-methylene diphophonate, removed bone metastatic lesions for cell primary culture, chromosome analysis for determine the bone metastatic cells have a characterization of unchanged humanization, in the anesthesia death mice. Through eight in vivo ~ in vitro selections, the 4th, 8th generation cells of SPC-A-1, 8th generation cells of NCI-H460 and their parental cells were used for microarray analysis, respectively.
Project description:Expression profile of parental wild type non-small cell lung cancer, NCI-H460, and cancer stem cell-rich (CSC-rich) populations treated with PNAs-A15 for 6 h. Results provide the information that PNAs-A15, a peptide nucleic acid of A-repeats length 15 bp, suppressed up-regulated A-repeats containing genes in both parental wild type and CSC-rich cells. In this study, we isolate cancer stem cell-rich (CSC-rich) population from a non-small cell lung cancer (NSCLC) cell line, NCI-H460, by selectively propagating the cells in a spheroid culture condition. The parental wild type H460 and CSC-rich cells were maintained at 37°C in 5% CO2 under sterile conditions in Roswell Park Memorial Institute (RPMI) 1640 medium. Cells were treated with PNAs-A15 in 6-well microplates for 6 h. Total RNA was isolated and hybridized on the HumanHT-12 v4 Expression BeadChip. The RNA expressions were evaluated.
Project description:The FHIT gene, at 3p14.2, encompasses the common fragile site FRA3B and is frequently inactivated in primary tumors and cell lines of lung, head and neck, stomach, esophagus, cervix and breast cancer. In particular, loss of the FHIT protein is one of the most frequent alteration in lung tumors and pre-invasive lung lesions, suggesting a role for this gene in the early stages of lung carcinogenesis. Adenoviral-mediated restoration of FHIT expression in FHIT-negative cell lines results in cell cycle alteration, induction of apoptosis (through activation of the cytoplasmic apoptotic pathway) and regression of the tumorigenic phenotype. Taken together these observations support the hypothesis of a role for FHIT in human carcinogenesis, but little is known about its mechanism of action. Identification of transcriptional targets of FHIT is therefore critical to understand the pathways by which FHIT promotes growth arrest and apoptosis.
Project description:Human lung adenocarcinoma cell lines (CPA-Yang1, CPA-Yang2, CPA-Yang3, SPC-A-1) and human larger cell lung cancer cell line NCI-H460 were injected into left cardiac ventricle of nude mice for bone metastases clone, respectively. Bone metastatic lesions were detected by bone scintigraphy with 99mTc-methylene diphophonate and X ray. Removed bone metastatic lesions for cell primary culture, chromosome analysis for determine the bone metastatic cells have a characterization of unchanged humanization, in the anesthesia death mice. Through in vivo ~ in vitro selections, the bone-seeking clones 1st, 3th, 4th, 8th, 8th passage cells of CPA-Yang1, CPA-Yang2, CPA-Yang3, SPC-A-1, NCI-H460 and their parent cells were used for microarray analysis, respectively.
Project description:The experiment was designed to identify the genes which get altered after the transfection of siRNA targeting STAT6 in NCI-H460 cells. The NCI-H460 cells plated in 12-well plate were transfected with 60nM of STAT6 specific siRNA. Biological duplicates of 2 samples were used viz. control NCI-H460 cells (Contol), NCI-H460 cells transfected with siRNA (siRNA)
Project description:The receptor tyrosine kinase (RTK) EGFR is overexpressed and mutated in NSCLC. These mutations can be targeted by RTK inhibitors (TKIs), such as erlotinib. Chromatin-modifying agents offer a novel therapy approach by sensitizing tumor cells to TKIs. The NSCLC cell lines HCC827 (EGFR mutant, adenocarcinoma), A549 (EGFR wt, adenocarcinoma) and NCI-H460 (EGFR wt, large cell carcinoma) were analyzed by SNP6.0 array. Changes in proliferation were quantified by WST-1 assay, apoptosis by Annexin V/7-AAD flow cytometry and histone marks (acH3, H3K4me1,-2,-3) by immunoblotting. Expectedly, the EGFR wt cell lines A549 and NCI-H460 were insensitive to the growth-inhibiting effect of single-agent erlotinib (IC50 70-100µM), compared to HCC827 (IC50 <0.02μM). Treatment with panobinostat diminished growth to <50% in both EGFR wt and <30% in HCC827 cells. The combination of both drugs significantly reduced proliferation by ≥70% in A549, >95% in HCC827, but not further in NCI-H460. Panobinostat alone induced differentiation and expression of p21WAF1/CIP1 and p53 in all three cell lines, with almost no further increase when combined with erlotinib. In contrast, combination treatment additively decreased pERK, pAKT and pEGFR in A549, and synergistically induced acH3 in both adenocarcinoma lines. Surprisingly, we also saw an induction of H3K4 methylation marks in all three cell lines. In conclusion, panobinostat synergistically sensitized lung adenocarcinoma cells to the antiproliferative effects of erlotinib. Since single-agent erlotinib has only modest clinical effects in adenocarcinoma EGFR wt patients, combination therapy with an HDACi might offer a promising therapy approach to extend this activity. Copy-number analysis of three NSCLC cell lines HCC827, A549 and NCI-H460 (in unicates) was performed according to protocol by Affymetrix Genome-Wide Human SNP-Array 6.0.
