A1e inhibits proliferation and induces apoptosis in nci. A, adriamycin adr inhibited the proliferation of ncih460 and a549 cells as measured by mtt assays. Thymoquinone and cisplatin as a therapeutic combination in. Bmp4smad signaling pathway mediates adriamycininduced.
Description luciferase stably expressing cell line ncih460 human nonsmall cell lung cancer also known as species human tissue growth properties morphology epitheliallike growth medium rpmi. Transfection of nih3t3 cells, hela, swis 3t3, 293t with lipofectamine 2000 1. Grand island, ny, usa, according to the manufacturers instructions. Thymoquinone tq is a compound extracted from black caraway seeds of nigella sativa and is active against various cancers. A1e inhibits proliferation and induces apoptosis in ncih460 lung cancer cells via extrinsic and intrinsic pathways. The side population in human lung cancer cell line nci. It can be used for transfection of adherent or suspension cells with dna or rna including stealth rnai and. Each reaction mix is sufficient for triplicate 96well, duplicate 24well, and single well 6well transfections, and accounts for pipetting variations. Ncih460 cells labeled with fluorescent nanoparticles fnp or green. For gfp gene transfection in the ncih460 cells, the lipofectamine 2000 invitrogen transfection was carried out following the general protocol invitrogen lipofectamine 2000 technical manual, see the. For gfp gene transfection in the ncih460 cells, the lipofectamine 2000 invitrogen transfection was carried out following the general protocol invitrogen. This tumorigenic cell line exhibits epithelial morphology and has been known to express p53 mrna as normal lung tissue does. Follow these important guidelines when transfecting ncih460 cells using.
Ncih460 reverse transfection protocol 24well plate. All cell lines were transfected using lipofectamine 2000 invitrogen in 24well. More the cosmic search system is a simple text search tool. I use lipofectamine 2000 invitrogen for transfection of both sirna and plasmid dna into these 293 cell lines. Dmc induces cell morphological changes and decreases the percentage of viable ncih460 cells.
Theres a difference between the two protocols lipofectamine 3000 has an extra reagent that needs to be added. Lipofectamine 2000 is a lipid based transfection reagent for eukaryotic cells. Nci h460, ncih520, ncih596, the human immortalized bronchial epithelial. This reference provides a recommended procedure to transfect plasmid dna into nci h460, human large cell lung cancer cells atcc no. Pdf primo vessel as a novel cancer cell migration path. Your search term ncih460 returned results in 1 section of the database. Jcrb luciferase expressing cancer cells ncih460luc. Transienttransfectionofa549,hct116,andncih460 cells were performed using the lipofectamine.
Establishment and characterization of a human lung cancer cell line ncih460lnm35 with consistent lymphogenous metastasis via both. Researchers use lipofectamine 2000 reagent for sirna and shrnabased gene knockdown experiments, as well as for gene expression studies. Ncih460, ncih358 and a549 cells were transfected, using. Optimemin ul, equal volume for dna and lipofectamine lipofectaminein ul 25.
Luteolin exerts an anticancer effect on ncih460 human nonsmall. Although the egf receptor tyrosine kinase inhibitors egfrtki gefitinib have shown dramatic effects against egfr mutant lung cancer, patients become resistant by various mechanisms. Lipofectamine 2000 transfection reagent is a proprietary formulation for the transfection of nucleic acids dna and rna into eukaryotic cells and provides the following advantages. The distant metastasis of cancer cells is a risk factor for tumor lethality and poor prognosis in nonsmallcell lung carcinoma nsclc. Primo vessel as a novel cancer cell migration path from. Lipofectamine 3000 transfection reagent leverages our most advanced lipid nanoparticle technology to provide superior transfection performance with improved application outcomes and reproducible. Protocol 6 well dish plate cells and grow to 70% confluence in 2 ml media without antibiotics dmem with 10% fbs. Lipofectamine 2000 reagent protocol 202 lipofectamine 2000 dna transfection reagent protocol transfect cells according to the following chart. Scaling up or down lipofectamine 3000 reagent transfections.
To atcc valued customers, atcc stands ready to support. Nci h460 standard transfection protocol 24well plate. Research articleprimo vessel as a novel cancer cell migration path from testis with nanoparticlelabeled and gfp expressing cancer cells hyunjung han1, hongbae kim1, jinmyung cha2, jinkyu. The ncih460 cells were treated with 0, 15, 20, 25, 30 and 35. Similarly, the fg30sirt1 or fg30 packaging plasmid and envelope protein. Lipofectamine 2000 transfection reagent from invitrogen. Lipofectamine 2000 transfection for hek293 cells about 6070% transfection rate transfect cells 6well plate. Ncih460 and a549 cells were treated with 500 n m and 2. Gibco rpmi 1640 with glutamax supplement 61870036 10% gibco fbs a3160401 0. Establishment and characterization of a human lung cancer. This tumorigenic cell line exhibits epithelial morphology and has been known to express. The lipofectamine 3000 is the bees knees though, i managed to get loads of transfects from. Luteolin exerts an anticancer effect on nci h460 human nonsmall cell lung cancer cells through the induction of sirt1mediated apoptosis.
Lipofectamine transfection of c2c12 cells bridges lab. Cy3 and cy5 dye following the protocol given in the ettan dige user. Transfection protocol lipofectamine 3000 transfection reagent nci h460 lung cancer cells complete growth medium component cat. Cisplatin cddp is the most active chemotherapeutic agent in lung cancer. Bd diagnostics systems, sparks, md, usa was used to isolate only gfpexpressedcells from the cultured cells. A novel naphthalimide compound restores p53 function in non. Polyplustransfections strengths are 20 years of experience in manufacturing transfection solutions with tailored scientific and regulatory support to accelerate research. Primo vessel as a novel cancer cell migration path from testis with. The nci h460 cell line is also unique in the fact that it shows no significant structural dna abnormalities. The cells express easily detectable p53 mrna at levels. Lipofectamine 2000 reagent thermo fisher scientific. Demethoxycurcumin induces the apoptosis of human lung.
Antitumor activity of selective mek12 inhibitor azd6244. The ncih460 human lung carcinoma cell line and hek293t cell line were. Transfection protocol of lipofectamine rnaimax can be. Invitrogen lipofectamine 3000 transfection reagent 1. Lipofectamine ltx reagent is a proprietary, animalorigin free formulation for the transfection of dna into eukaryotic cells with low cytotoxicity. Transfection of nih3t3 cells, hela, swis 3t3, 293t with. Lipofectamine 2000 transfection reagent is versatile transfection reagent that has been shown to effectively transfect widest variety of adherent and suspension cell lines. Our first job is to listen to and observe what our customers need, and meet those needs. Luteolin exerts an anticancer effect on ncih460 human non. Ncih460, hct116, and a549 cells were maintained in. Transfection protocol lipofectamine 3000 transfection. Ncih460 cells were seeded into 6well plates 3 105 cells well or 24well. Usa were transfected using lipofectamine 2000 invitrogen. Nci h460 reverse transfection protocol 24well plate.
To atcc valued customers, atcc stands ready to support our customers needs during the coronavirus pandemic. Transfecting plasmid dna into nci h460 cells using lipofectamine 3000 reagent. Ncih460 standard transfection protocol 24well plate. Recent studies indicate that tumors contain a subset of stemlike cancer cells that possess certain stem cell properties. Lipofectamine 2000 transfection reagent dartmouth college. Brightfield image of nci h460 cells in culture, prior to transfection. Herein, we used hoechst 33342 dye efflux assay and flow. For gfp gene transfection in the ncih460 cells, the lipofectamine 2000 invitrogen transfection was carried out following the general protocol invitrogen lipofectamine 2000 technical manual, see the supporting information for our particular procedure and a previous study. The cells show a bright green fluorescence evenly distributed throughout the cytosol. Damtc regulates cytoskeletal reorganization and cell. Effi ciency and ofp expression were analyzed 72 hours. Piperlongumine inhibits lung tumor growth via inhibition. Stat6 silencing upregulates cholesterol synthesis via mir.
Primo vessel as a novel cancer cell migration path from testis with nanoparticlelabeled and gfp expressing cancer cells. Ncih460 transfection kit for lung carcinoma cells, htb177. H460 ncih460 nonsmall human lung carcinoma cells were stably transfected with a plasmid coding for turbogfp. Download scientific diagram ncih460, ncih358 and a549 cells were transfected, using lipofectamine 2000, with bc819 and cotransfected with a plasmid. Invitrogen lipofectamine 2000 transfection reagent. Furthermore, overexpression of mir197 after stat6 sirna transfection in ncih460 cells reversed the effect of stat6 silencing on cholesterol levels fig. North american service manual model pc series version 2 march 25, 1997 revision b1. Transfecting plasmid dna into ncih460 cells using lipofectamine. Gazdar and colleagues in 1982 from a male with large cell lung carcinoma.