Abstract
BackgroundGastric cancer is the second most common cause of cancer-related death in males and the fourth in females. Traditional treatment has poor prognosis because of recurrence and systemic side effects. Therefore, the development of new therapeutic strategies is an important issue. Lentivirus-mediated shRNA stably inhibits target genes and can efficiently transduce most cells. Since overexpressed cyclin D1 is closely related to human gastric cancer progression, inhibition of cyclin D1 using specific targeting could be an effective treatment method of human gastric cancer.MethodsThe therapeutic effect of lentivirus-mediated shRNA targeting of cyclin D1 (ShCCND1) was analyzed both in vitro and in vivo experiments.ResultsIn vitro, NCI-N87 cells with downregulation of cyclin D1 by ShCCND1 showed significant inhibition of cell proliferation, cell motility, and clonogenicity. Downregulation of cyclin D1 in NCI-N87 cells also resulted in significantly increased G1 arrest and apoptosis. In vivo, stable NCI-N87 cells expressing ShCCND1 were engrafted into nude mice. Then, the cancer-growth inhibition effect of lentivirus was confirmed. To assess lentivirus including ShCCND1 as a therapeutic agent, intratumoral injection was conducted. Tumor growth of the lentivirus-treated group was significantly inhibited compared to growth of the control group. These results are in accordance with the in vitro data and lend support to the mitotic figure count and apoptosis analysis of the tumor mass.ConclusionThe lentivirus-mediated ShCCND1 was constructed, which effectively inhibited growth of NCI-N87-derived cancer both in vitro and in vivo. The efficiency of shRNA knockdown and variation in the degree of inhibition is mediated by different shRNA sequences and cancer cell lines. These experimental results suggest the possibility of developing new gastric cancer therapies using lentivirus-mediated shRNA.
Highlights
Gastric cancer is the second most common cause of cancer-related death in males and the fourth in females
shRNA targeting of cyclin D1 (ShCCND1) significantly inhibited cyclin D1 expression and cell proliferation in NCI-N87 cells NCI-N87 cells infected with lentivirus were successfully selected in medium containing puromycin (1 μg/ml)
The expression level of pRB, a downstream molecule of cyclin D1, showed significant inhibition in ShCCND1. These data suggest that lentivirus-mediated ShCCND1 could effectively inhibit endogenous cyclin D1 expression in NCI-N87 cells
Summary
Gastric cancer is the second most common cause of cancer-related death in males and the fourth in females. Traditional treatment has poor prognosis because of recurrence and systemic side effects. Since overexpressed cyclin D1 is closely related to human gastric cancer progression, inhibition of cyclin D1 using specific targeting could be an effective treatment method of human gastric cancer. Gastric cancer is the one of the most common causes of cancer-related death all over the world. The survival of gastric cancer patients has improved owing to improvement in diagnostic and therapeutic tools [1]. Cyclin D1, a cell cycle regulator, is closely related to several human cancers. Inhibition of cyclin D1 may be an innovative strategy for cancer treatment [4,5]
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