The global gene-expression profiles of U-937 human macrophages treated with Tat peptide and Tat-FITC conjugate.
There is increased interest in using cell penetrating peptides such as HIV Tat-derived peptide (Tat) to deliver biologically active cargo to cells. However, little is known about the precise molecular mechanism in cells after the uptake of Tat and after it conjugates with the cargo. Using high-density microarray analysis, we evaluated the alteration of whole genomic responses in U-937 macrophages that had been treated with Tat and Tat-fluorescein-5-isothiocynate (FITC), which served as model cargo, and then incubated for 24 h. Sixteen genes were upregulated and 28 downregulated in Tat-treated cells. After FITC had conjugated to Tat, 13 genes were upregulated and 23 downregulated. Ten upregulated and 13 downregulated genes were overlapped by both Tat and Tat-FITC. The molecular functions of regulated genes affected by Tat and Tat-FITC are described. A real-time quantitative reverse transcriptase-polymerase chain reaction was used to confirm three regulated genes (IFNAR2, CASP8, and CRLF1) affected by both Tat and Tat-FITC. We demonstrated that regulating the three genes was time-dependent and that CASP8 is also kinetically regulated in HeLa cells. Understanding the influences and consequences of Tat (Tat-cargo)-induced gene changes should help us design and develop efficient and safe delivery systems into target cells.
Institute of Microbiology and Immunology, Division of Allergy, Immunology, and Rheumatology, Medical College of Chung Shan Medical University, Taichung, Taiwan.
Chia-Wei
CW
Lin
Jung-hua Steven
JH
Kuo
Ming-shiou
MS
Jan
D053181:Caspase 8 / Q000235:genetics; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers / Q000737:chemistry / Q000494:pharmacology*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000737:chemistry / Q000494:pharmacology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D046228:Microarray Analysis; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D053633:Receptor, Interferon alpha-beta / Q000235:genetics; D018121:Receptors, Cytokine / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020298:U937 Cells; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry / Q000494:pharmacology*
D057846:Cell-Penetrating Peptides; D004337:Drug Carriers; D005452:Fluoresceins; C508033:IFNAR2 protein, human; D010446:Peptide Fragments; D018121:Receptors, Cytokine; C575807:Tat peptide (47-59), HIV-1; C113554:cytokine-like factor-1; D054322:tat Gene Products, Human Immunodeficiency Virus; D053633:Receptor, Interferon alpha-beta; C505530:CASP8 protein, human; D053181:Caspase 8