10.3748\/wjg.v5.i1.18<\/a><\/p>\nAuthors:<\/h4>\n
Bing Chen<\/p>\n
Abstract:<\/h4>\n
To further investigate the effect of cyclin D1 on the bio logic behavior of cancer cells and its potential role in gene therapy of tumor. METHODS A cyclin D1 subcloning plasmid termed BKSD1 was const ructed by subcloning the human cyclin D1 cDNA into Bluescript-KS, a plasmid vec tor with a pair of T7 and T3-promoters, with recombinant DNA technology of molecular biology. So, it is easy to generate digoxigenin (DIG)-labeled RNA probes of antisense and sense to cyclin D1 using RKSD1 as a template vector. PDORD1AS, an eukaryotic expression vector containing the full-length human cyclin D1 cDNA in its antisense orientation cloned into the retroviral vector pDOR-neo, was succ essfully constructed with BKSD1 to change restriction sites. A gastric cancer cell line, SGC7901\/VCR, was transfected with pDORD1AS by Lipofect Amine-mediate d introduction and a subline termed SGC7901\/VCRD1AS, which had stable overexpr ession of antisense RNA to cyclin D1, was obtained by selection in G418. The sub line, control subline transfected pDOR-neo and SGC7901\/VCR were evaluated by methods of immunohistochemistry, flow cytometry, molecular hybridization, morpho logy and cell biology. RESULTS Compared with control cell lines, SGC7901\/VCRD1AS had a reduced expression of cyclin D1 (inhibition rate was about 36%), in-creased ce ll size and cytoplasm to nucleus ratio, increased doubling time (42.2 h to 26.8 h and 26.4 h), decreased saturation density (18.910 4 to 4.810 5 and 4.810 5 ), increased percentage of cells in the G1\/G0 phase (80.9%-64.6% and 63.8%), reacquired serum dependence, and a loss of tumorigenici ty in nude mice (0\/4 to 4\/4 and 4\/4). CONCLUSION Stable overexpression of antisense RNA to cyclin D1 can reverse the transformed phenotype of human gastric cancer cells and may provide an approach of gene therapy for gastric cancer.<\/p>\n","protected":false},"excerpt":{"rendered":"
DOI: 10.3748\/wjg.v5.i1.18 Authors: Bing Chen Abstract: To further investigate the effect of cyclin D1 on the bio logic behavior of cancer cells and its potential role in gene therapy of tumor. METHODS A cyclin D1 subcloning plasmid termed BKSD1 was const ructed by subcloning the human cyclin D1 cDNA into Bluescript-KS, a plasmid vec tor […]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[9],"tags":[],"class_list":["post-390","post","type-post","status-publish","format-standard","hentry","category-its-e"],"_links":{"self":[{"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/posts\/390","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=390"}],"version-history":[{"count":0,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/posts\/390\/revisions"}],"wp:attachment":[{"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=390"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=390"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=390"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}