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Inhibiting effect of miR-486-5p on xenograft of gastric cancer in nude mice
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Abstract:
Objective:To explore the effect of miR-486-5p on gastric cancer xenograft in nude mice and to investigate its probable mechanism. Methods: The subcutaneously transplanted tumor models of human gastric carcinoma (SGC-7901 cell line) in nude mice were established and after treated with miR-486-5p over-expression plasmids, growth situation of cancer xenografts in the nude mice was observed. The expression of NRP2（the target gene of miR-486-5p）were detected by Western blotting and immunohistochemical method. Results：Gastric carcinoma (SGC-7901) xenograft models in nude mice were successfully constructed; miR-486-5p can significantly inhibit the growth of xenografts in nude mice and the expression of miR-486-5p in cancer xenografts of experimental group was significantly higher than that of control group (P＜0.05). Compared to the negative control group and blank control group, the average mass and volume of cancer xenografts in experiment group was significantly less than those in the other groups, mass:(0.404±0.080) g vs (0.748±0.122) g, (0.788±0.176) g, all P＜0.05; volume: (0.333±0.039) cm3 vs (0.597±0.175) cm3, (0.594±0.216) cm3, P＜005, and the inhibition rate of cancer xenografts in experiment group was 46.99%. After treated with miR-486-5p over-expression plasmid in experiment group, the immunohistochemical results showed that NRP2 protein, presenting as yellow particle, mainly existed in the cytoplasm of gastric cancer cells; the IHS score of NRP2 protein in miR-486-5p group was significantly lower than that of NC and blank control groups (\[2.2±0.84\] vs \[6.4±0.89\], \[6.2±1.48\], all P<001); however, there was no significant difference between negative control and blank control groups (P＞005). The results of Western blotting shown that the relative expression of NRP2 protein in cancer xenograft tissues of experiment group was significantly less than that of the other groups (\[0.04±0.006\] vs \[0.70±0.03\], (0.68±002\]，all P<001). The difference in IHS score, inhibition rate of cancer and NRP2 protein expression between NC and blank control groups had no statistical significance (P>0.05). Conclusion: miR-486-5p can remarkably inhibit the growth of gastric cancer xenograft in nude mice. It may associate with inhibiting expression of NRP2.
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Project Supported:
Project supported by the Natural Science Foundation of the Shandong Province （No. ZR2014HQ072），the Science and Technology Development Program in Shandong Province（No. 2014WS0490），and the Science and Technology Development Program in Binzhou City（No. 2014ZC0116）
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