title: Effect of shRNA targeting mouse CD99L2 gene in a murine B cell lymphoma             in vitro and in vivo.
abstract: Mouse CD99 antigen-like 2 (mCD99L2) has previously been confirmed to be             expressed in murine B lymphoma (A20) cells by our group. The present study aimed             to establish a mCD99L2‑downregulated A20 cell line and to investigate the effect             of shRNA targeting mCD99L2 in A20 cells in vitro and in vivo. Four pLenti6/mCD99L2             expression vectors containing the mCD99L2 shRNA-expressing cassette were constructed,             transfected into A20 cells and stable mCD99L2-downregulated A20 subclones, termed             A20-mCD99L2- cells, were established and identified by quantitative PCR and western             blot analysis. Light and transmission electron microscopy, MTT assay, flow cytometry             and immunofluorenscence labeling were used to observe the morphological, biological             and phenotypic characteristics in vitro. Some of the A20-mCD99L2- cells exhibited             H/RS‑cell like morphology, a decreased proliferative ability, a prolonged G2 phase             and increased CD30 and CD15 expression. Upon injecting cells into nude or immunocompetent             BALB/c mice, tumorigenesis, tumor growth, morphology and phenotypes in vivo were             observed. A20-mCD99L2- cells induced tumors in nude and BALB/c mice, but with             less potency in the latter compared with the controls. Similar morphological,             biological and phenotypic characteristics were observed in the A20-mCD99L2- cell-induced             tumors as those in vitro. Several cytokines including CD30T, IL-12p40/p70, IL-3,             IFN-γ, CXCL16, MIP-1α and CD40 were upregulated following mCD99L2 downregulation             when detected using antibody arrays. The results from western blot analysis indicated             that the regulation of mCD99L2 expression may involve the activated nuclear factor-κB             pathway in the murine B lymphoma cells. The present study provides data for further             investigation into the mCD99L2 gene in tumor cells.
authors: Department of Pathology, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, PR China.
authors: Fang
authors: F
authors: Liu
authors: Gong
authors: G
authors: Zhang
authors: Fanrong
authors: F
authors: Liu
authors: Xinhua
authors: X
authors: Zhou
authors: Xiaoyan
authors: X
authors: Chen
authors: Xiqun
authors: X
authors: Han
authors: Ziqing
authors: Z
authors: Wu
authors: Tong
authors: T
authors: Zhao
mesh_terms: D000071443:12E7 Antigen; D000818:Animals; D015703:Antigens, CD / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D054405:Chemokine CCL3 / Q000235:genetics; D006801:Humans; D007377:Interleukin-3 / Q000235:genetics; D016393:Lymphoma, B-Cell / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
chemical_list: D000071443:12E7 Antigen; D015703:Antigens, CD; C522605:Cd99 protein, mouse; D054405:Chemokine CCL3; D007377:Interleukin-3; D034741:RNA, Small Interfering