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: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.
Project description:Background: Leiomyosarcomas are among the most common histological types of soft tissue sarcoma (STS), with no effective treatment available for advanced patients. Lung metastasis, the most common site of distant metastasis, is the primary prognostic factor. We analysed the immune environment targeting lung metastasis of STS to explore new targets for immunotherapy. Methods: We analysed the immune environment of primary and lung metastases in 38 patients with STS using immunohistochemistry. Next, we performed gene expression analyses on primary and lung metastatic tissues from six patients with leiomyosarcoma. Using human leiomyosarcoma cell lines, the effects of the identified genes on immune cells were assessed in vitro. Results: Immunohistochemistry showed a significant decrease in CD8⁺ cells in the lung metastases of leiomyosarcoma. Among the genes upregulated in lung metastases, epithelial cellular adhesion molecule (EPCAM) showed the strongest negative correlation with the number of CD8⁺ cells. Transwell assay results showed that the migration of CD8⁺ T cells was significantly increased in the conditioned media obtained after inhibition or knock down of EPCAM. Conclusions: EPCAM was upregulated in lung metastases of leiomyosarcoma, suggesting inhibition of CD8⁺ T cell migration. Our findings suggest that EPCAM could serve as a potential novel therapeutic target for leiomyosarcoma.
Project description:Targeted therapies against cancer stem cells which are enriched in side populations (SP) involves interruption of Wnt-signalling. Furthermore, EpCAM is a SP marker and modulator of Wnt-signalling. Therefore, the effects of an anti-EpCAM treatment on SP-cells and WNT/β-catenin signalling was studied. SP of the murine lung adenocarcinoma cell line A2C12 was obtained by fluorescence activated cell sorting and whole genome scans helped to define their molecular phenotype after anti-EpCAM antibody treatment.