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Archive > Volume 20 Issue 3 > 2013,20(3):312-316. DOI:10.3872/j.issn.1007-385X.2013.03.010 Prev Next
Basic Research
Adenovirus mediated over-expression of miRNA-29a inhibits proliferation of human gastric cancer cells
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Abstract:
Objective:To explore the inhibitory effect of miRNA-29a(miR-29a) on the proliferation of human gastric cancer cell lines SGC-7901 and AGS through over-expression of miR-29a mediated by the recombinant replication-deficient human adenovirus type 5 vector. Methods: The recombinant adenovirus Ad-miR-29a containing pre-miR-29a or control adenovirus Ad-LacZ containing LacZ gene was constructed and infected into human gastric cancer SGC-7901 and AGS cells, respectively. The expressions of miR-29a in SGC-7901 and AGS cells were detected by real-time PCR. CCK-8 assay was employed to examine the inhibitory effect of miR-29a on the proliferation of human gastric cancer cell lines. The flow cytometry assay was used to analyze cell cycle of SGC-7901 and AGS cells. The migration ablilities of SGC-7901 and AGS cells were assessed by Transwell assay. Results: Compared with the Ad-LacZ group, the Ad-miR29a group expressed higher level of miR-29a in both SGC-7901 and AGS cells (\[17.35±0.71\] vs \[1.12±0.09\], \[26.50±109\] vs \[0.95±0.04\], P<001). The proliferation of SGC-7901 and AGS was significantly inhibited in the Ad-miR29a group as compared with that in the Ad-LacZ group. Six days after Ad-miR29a infection, the D value was significantly decreased (\[0.54± 003\] vs \[0.77 ± 0.04\], \[0.70 ± 0.03\] vs \[0.88 ± 0.04\], P<0.01). The proliferation of cells in the Ad-LacZ group showed no significant changes as compared with that in the uninfected group (P>005). Meanwhile, The percentage of SGC-7901 or AGS cells arrested in G0/G1 period in the Ad-miR29a group was significantly higher than that in the Ad-LacZ group (\[63.10±4.91\]% vs \[47.60±5.31\]%，\[69.80±3.15\]% vs \[5460±422\]%, P<0.05\]. However, no significant changes were found in the migration ability of gastric cancer SGC-7901 and AGS cells. Conclusion:miR-29a can effectively suppress the proliferation of human gastric cancer cells, which makes a promising new therapeutic target for gastric cancer.
Keywords:
miRNA-29a(miR-29a) ; adenovirus ; gastric cancer ; proliferation ; cell cycle
Project Supported:
Project supported by the Natural Science Foundation of Chongqing (No. CSTC,2010BB5158, No. CSTC, 2011AC5024, No. CSTC, 2011jjA0878, No. CSTC, 2011jjA0882)
