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Archive > Volume 20 Issue 6 > 2013,20(6):667-672. DOI:10.3872/j.issn.1007-385X.2013.06.006 Prev Next
Basic Research
Lentivirus mediated scilencing of EGFL7 gene and its effects on biological behavior of human lung carcinoma A549 cells in vitro
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Abstract:
Objective:To investigate the effect of epidermal growth factor-like domain 7 ( EGFL7 ) on the proliferation, apoptosis, invasion and angiogenesis of lung carcinoma A549 cells through silence EGFL7 by lentivirus infection. Methods: Lentivirus mediated small hairpin RNA target EGFL7 (LV-RNAi) were transfected into A549 cells. Three groups were designed in this study: a LV-RNAi group (A549 cells transfected with LV-RNAi), a LV-NC group (A549 cells transfected with the control GFP-lentivirus) and a control group (A549 cells). Real-time PCR and Western blotting were performed to detect the effect of LV-RNAi infection on the expression of EGFL7 mRNA and protein in A549 cells. MTT assay and flow cytometry were used to analyz the influence of silence EGFL7 on the growth and apoptosis of A549 cells, respectively. Transwell invasion assay and chick embryo chorioallantoic membrane (CAM) assay were used to detect the influence of silence EGFL7 on the angiogenesis and invasion, respectively. Results: Lentivirus infected A549 cells stably and efficiently. Expression of EGFL7 mRNA （\[0.18±0.03\] vs \[0.98±0.05\], \[1.03±0.09\]; P<0.05） and protein of the LV-RNAi group were significantly reduced compared with that of the LV-NC group and control group. Compared with the LV-NC group and control group, silencing EGFL7 showed no obvious influence on the proliferation capacity (120 h after infection: \[0.73±0.22\] vs \[0.79±0.08\] vs \[0.81±0.11\], P>0.05) and apoptosis rate (120 h after infection: \[1.92%±0.06\] vs \[2.11%±0.07\] vs \[1.85%±0.13\], P>0.05) of A549 cells in the LV-RNAi group; meanwhile, the invasion cell number (\[61.7±14.5\] vs \[272.8±21.5\], \[286.6±40.0\]/mm2; P<0.05) and the angiogenesis index (\[31.7±4.1\] vs \[96.3±4.4\], \[103.3±6.5\]; P<0.05) were significantly decreased in the LV-RNAi group. Conclusion: Silencing EGFL7gene may inhibit the invasion ability and the angiogenesis ability of A549 cells without affecting the proliferation and apoptosis.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No. 30972718）, the Natural Science Foundation of Jiangsu Province (No. BK2012606), the Post-graduate Culture and Innovation Foundation of Jiangsu Province（No. CXZZ12_0837）and the Natural Science Foundation of Colleges and Universities in Jiangsu Province（No. 13KJB320019）
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