Project description:We extracted RNA of sorted lung SPC/GFP+ EpCAM+ cells and performed microarray analyses. Total RNA was extracted from sorted Ep-CAMhigh/GFPhigh cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. The RNA integrity was examined on an Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA). Biotinylated ss-cDNA were prepared according to the standard Affymetrix protocol from 100 ng total RNA using the GeneChip WT PLUS Reagent Kit User Manual (Affymetrix/Thermo Fisher Scientific). Fragmented and labeled ss-cDNA were hybridized on a GeneChip Clariom S Array (Affymetrix/Thermo Fisher Scientific) (n =3/group). We used microarrays to detail the global gene expression of SPC/GFP+ EpCAM+ cells from the control mice(G29-5_(Clariom_S_Mouse)) and the 3 months smoke mice(G29-6_(Clariom_S_Mouse)), and identified distinct classes of up-regulated genes during this process.
Project description:We extracted RNA of sorted lung SPC/GFP+ EpCAM+ cells and performed microarray analyses. Total RNA was extracted from sorted Ep-CAMhigh/GFPhigh cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. The RNA integrity was examined on an Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA). Biotinylated ss-cDNA were prepared according to the standard Affymetrix protocol from 100 ng total RNA using the GeneChip WT PLUS Reagent Kit User Manual (Affymetrix/Thermo Fisher Scientific). Fragmented and labeled ss-cDNA were hybridized on a GeneChip Clariom S Array (Affymetrix/Thermo Fisher Scientific) (n =3/group). We used microarrays to detail the global gene expression of SPC/GFP+ EpCAM+ cells from the control mice(G37-1_(Clariom_S_Mouse)), 3 week continuous smoke mice (G37-2_(Clariom_S_Mouse)) and 3 weeks intermittent smoke mice(G37-3_(Clariom_S_Mouse)), and identified distinct classes of up-regulated genes during this process.
Project description:EpCAM is one of the highly expressed marker in esophageal adenocarcinoma as identidied in several multi-OMICS platform. We utilize FLO-1 cells which are lacking in EpCAM expresssion and use overexpression strategy to elucidate the function of EpCAM in this cells. Recombinant EpCAM-GFP or GFP only was trasnfected in FLO-1 cells, and the stable cells after antibiotic selection were isolated and expanded. The cells were then subjected to mRNA sequencing.
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: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:Healthy donor and idiopathic pulmonary fibrosis (IPF) patient lung tissues were digested, Lin–EpCAM+ cells were isolated with FACS, and then single-cell RNA-seq was performed.
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.