title: Noninvasive visualization of microRNA-16 in the chemoresistance of gastric cancer using a dual reporter gene imaging system.
abstract: MicroRNAs (miRNAs) have been implicated to play a central role in the development of drug resistance in a variety of malignancies. However, many studies were conducted at the in vitro level and could not provide the in vivo information on the functions of miRNAs in the anticancer drug resistance. Here, we introduced a dual reporter gene imaging system for noninvasively monitoring the kinetic expression of miRNA-16 during chemoresistance in gastric cancer both in vitro and in vivo. Human sodium iodide symporter (hNIS) and firefly luciferase (Fluc) genes were linked to form hNIS/Fluc double fusion reporter gene and then generate human gastric cancer cell line NF-3xmir16 and its multidrug resistance cell line NF-3xmir16/VCR. Radioiodide uptake and Fluc luminescence signals in vitro correlated well with viable cell numbers. The luciferase activities and radioiodide uptake in NF-3xmir16 cells were remarkably repressed by exogenous or endogenous miRNA-16. The NF-3xmir16/VCR cells showed a significant increase of (131)I uptake and luminescence intensity compared to NF-3xmir16 cells. The radioactivity from in vivo (99m)Tc-pertechnetate imaging and the intensity from bioluminescence imaging were also increased in NF-3xmir16/VCR compared with that in NF-3xmir16 tumor xenografts. Furthermore, using this reporter gene system, we found that etoposide (VP-16) and 5-fluorouracil (5-FU) activated miRNA-16 expression in vitro and in vivo, and the upregulation of miRNA-16 is p38MAPK dependent but NF-κB independent. This dual imaging reporter gene may be served as a novel tool for in vivo imaging of microRNAs in the chemoresistance of cancers, as well as for early detection and diagnosis in clinic.
authors.affiliation: School of Life Sciences and Technology, Xidian University, Xi'an, Shaanxi, China.
authors.forename: Fu
authors.initials: F
authors.lastname: Wang
authors.forename: Xinxing
authors.initials: X
authors.lastname: Song
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Xin
authors.forename: Shenxu
authors.initials: S
authors.lastname: Wang
authors.forename: Weidong
authors.initials: W
authors.lastname: Yang
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Kaichun
authors.initials: K
authors.lastname: Wu
authors.forename: Xiaoyuan
authors.initials: X
authors.lastname: Chen
authors.forename: Jimin
authors.initials: J
authors.lastname: Liang
authors.forename: Jie
authors.initials: J
authors.lastname: Tian
authors.forename: Feng
authors.initials: F
authors.lastname: Cao
mesh_terms: D000818:Animals; D002452:Cell Count; D045744:Cell Line, Tumor; D003952:Diagnostic Imaging; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D005260:Female; D005472:Fluorouracil / Q000494:pharmacology / Q000627:therapeutic use; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D017930:Genes, Reporter; D017405:Herpes Simplex Virus Protein Vmw65 / Q000378:metabolism; D006801:Humans; D049409:Luciferases, Firefly / Q000378:metabolism; D008163:Luminescent Measurements; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D013274:Stomach Neoplasms / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D027981:Symporters / Q000378:metabolism; D019076:Transgenes; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D000099209:Sodium-Iodide Symporters
chemical_list: D005472:Fluorouracil; D017405:Herpes Simplex Virus Protein Vmw65; D049409:Luciferases, Firefly; D035683:MicroRNAs; D016328:NF-kappa B; D027981:Symporters; D048051:p38 Mitogen-Activated Protein Kinases; D000099209:Sodium-Iodide Symporters; C505362:MIRN16 microRNA, human