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Archive > Volume 23 Issue 3 > 2016,23(3):335-342. DOI:10.3872/j.issn.1007-385X.2016.03.007 Prev Next
Basic Research
Influence of Ad.RGD-ING4-PTEN on proliferation, apoptosis and invasion of neuroglioma U87 cell
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Abstract:
Objective:To explore effect of the adenovirus vector (Ad.RGD-ING4-PTEN) modified with RGD and co-expressing with double genes coding inhibitor of growth 4 (ING4) and phosphatase and tensin homologue deleted on chromosome ten (PTEN) on proliferation, apoptosis and invasion of human neuroglioma cells in vitro.Methods: Neuroglioma U87 cells were in vitro infected with Ad.RGD-ING4-PTEN as experimental group, Ad.RGD-ING4/-PTEN as monogenge control group and PBS, Ad.RGD/RGD-GPF as blank control group. Expressions of the target genes, ING4 and PTEN in the U87 cells were detected by Western blotting. Effect of infection with the Ad.RGD-ING4-PTEN of experimental group on proliferation of the U87 cell was detected by MTT assay. Changes in apoptosis of the neuroglioma cells and expressions of apoptosis-related genes, Bcl-2, Bax, caspase-3 and HIF-1α, were respectively examined with flow cytometry and Real-time PCR assays. Effects of infection with the Ad.RGD-ING4-PTEN of experimental group on migration and invasion abilities were detected with scratch and Transwell assays respectively. Expression of invasion-associated genes, MMP-2 and MMP-9, was detected by Real-time PCR. Results: Expressions of ING4 and PTEN were successfully detected only in the experimental group and the corresponding monogene control groups. Inhibitory and apoptosis rate of the U87 cell on 5th day in experimental group were (83.1±4.6)% and (40.7±4.3)%, respectively, compared to the blank control and monogene control groups, the differences were statistically significant (P<0.05). In the experimental group, expressions of apoptosis-related proteins, Bax and caspase-3, were remarkably up-regulated, while expressions of HIF-1α and Bcl-2 proteins down-regulated in the U87 cells (all P<0.05), and expressions of invasion-associated molecules MMP-2 and MMP-9 also down-regulated significantly (all P<0.05 ). Migration distances(\[70.1±6.2\] μm) of the cells and number of the penetrated cells (\[266±3.5\] cell) in the experiment group were significantly lesser than those in the monogene control and blank control groups(all P<0.05). Conclusion: The adenovirus with double-gene ING4 and PTEN (Ad.RGD-ING4-PTEN) could more significantly inhibit proliferation and induce apoptosis of the neuroglioma U87 cells, compared to those in the adenovirus with monogene, and also could inhibit migration and invasion abilities of the U87 cells.
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Project Supported:
Project supported by the National Natural Science Foundation for Young Scholars of China (No. 81001016)
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