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Archive > Volume 20 Issue 2 > 2013,20(2):207-211. DOI:10.3872/j.issn.1007-385X.2013.02.014 Prev Next
Basic Research
KLK7 siRNA on gastric cancer AGS cell lines
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Abstract:
Objective: Four siRNA fragments targeting kallikrein-related peptidase 7 ( KLK7 ) were synthesized in vitro. The most effective siRNA was selected, and the effect of silencing KLK7 expression on proliferation and apoptosis of gastric carcinoma AGS cells was observed. Methods: Four siRNA fragments targeting KLK7 (KLK7-siRNA-416, KLK7-siRNA-596, KLK7-siRNA-474 and KLK7-siRNA-795) were designed and transiently transfected into AGS cells. qRT-PCR was used to detect the expression of KLK7 mRNA in each interference group. Western blotting was used to detect the protein expression of HK7 (encoded by KLK7 gene). AGS cell proliferation, and the cells cycle and apoptosis after transfection were detected by MTT assay and flow cytometry, respectively. Results: Among four KLK7-siRNAs, KLK7-siRNA-416 showed the highest interference efficiency. The ratio of KLK7 mRNA expression in KLK7-siRNA-416 group was significantly lower than those in control group (\[0.32±0.049)% vs \[0.93±0.071\], P<0.01). The protein expression of HK7 in KLK7-siRNA-416 group after transfection for 48 h was significantly decreased (\[1.18±0.198\] vs \[0.52±0096\], P<0.01). The cell proliferation of KLK7-siRNA-416 group was significantly inhibited after transfection for 48 h, with inhibition rate of (37.70±0.12)%, P<0.05. Cell cycles were blocked in G0/G1 phase by transfection. However, no impact was found in AGS cell apoptosis. Conclusion: The silencing expression of KLK7 by KLK7-siRNA inhibited the AGS cell proliferation and block cell cycels in G0/G1 phase. However, no impact was found in AGS cell apoptosis.
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