科创中国

Antisense to cyclin D1 reverses the transformed phenotype of human gastric cancer cells

• 摘要：

  AIM: To further investigate the effect of cyclin D1 on the biologic behavior of cancer cells and its potential role in gene therapy of tumor. METHODS: A cyclin D1 subcloning plasmid termed BKSD1 was constructed by subcloning the human cyclin D1 cDNA into Bluescript-KS, a plasmid vector 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 successfully constructed with BKSD1 to change restriction sites. A gastric cancer cell line, SGC7901/ VCR, was transfected with pDORD1AS by Lipofect Amine mediated introduction and a subline termed SGC7901/ VCRD1AS, which had stable overexpression of antisense RNA to cyclin D1, was obtained by selection in G418. The subline, control subline transfected pDOR-neo and SGC7901/ VCR were evaluated by methods of immunohistochemistry, flow cytometry, molecular hybridization, morphology and cell biology. RESULTS: Compared with control cell lines, SGC7901/ VCRD1AS had a reduced expression of cyclin D1 (inhibition rate was about 36%), increased cell size and cytoplasm to nucleus ratio, increased doubling time (42.2 h to 26.8 h and 26.4 h), decreased saturation density (18.9 × 10⁴ to 4.8 × 10⁵ and 4.8 × 10⁵), increased percentage of cells in the G1/ G0 phase (80.9% - 64.6% and 63.8%), reacquired serum dependence, and a loss of tumorigenicity 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.

• 作者：

  Bing Chen;Xue Yong Zhang;Yu Jing Zhang;Ping Zhou;Yan Gu;代明 樊

• 刊名：

  World Journal of Gastroenterology

• 在线出版时间：

  1999-2