Effect of lentiviral vector mediated RNA binding protein quaking-5 on the proliferation and apoptosis of lung adenocarcinoma cell line H1299

doi:10.16098/j.issn.0529-1356.2020.01.007

Abstract

Abstract:

To investigate the proliferation and apoptosis of lung adenocarcinoma cells line H1299 by the lentiviral vector mediated RNA binding protein quaking-5 (QKI-5). Methods GV358 (up-regulation) and GV248 (down-regulation) vectors were used to construct the lentiviral QKI-5 up-regulation vector and down-regulation vector, respectively. The vectors were transfected into 293T cells for lentiviral packaging and viral titer were then determined. Gene sequencing was performed to screen the sequence of vectors. Then Real-time PCR was used to evaluate the expression of QKI-5 mRNA and the proliferation of H1299 cells was examined by colony forming assay after transfection. The apoptosis of H1299 cells was determined by the detection of the expression membrane protein Ⅴ (annexin Ⅴ) and propyl iodide ingot (PI) by flow cytometry. Pro-apoptotic protein Caspae-3 and Caspase-8 were evaluated by Western blotting. Results QKI-5 up-regulation and down-regulation lentiviral vectors were constructed successfully. Compared with the controls, the expression of QKI-5 mRNA of H1299 cells was up-regulated, the cell colony formation was decreased, and the early apoptosis of H1299 cells was increased with the over-expression of Caspase-8 after transfected with up-regulated vector, whereas transfecting with QKI-5 down-regulated vector had opposite effect. Conclusion Lenviral vector mediated QKI-5 could inhibit proliferation and promote apoptosis of lung adenocarcinoma cells through Caspase-8.

Key words: Lung adenocarcinoma, RNA binding protein quaking quaking-5, H1299 cell line, Vector construction, Lentiviral transfeted, Real-time PCR

CLC Number:

R730.7

FANG Ke-xin WANG Xin-chen HONG Kai-ting ZHU Wang-yu. Effect of lentiviral vector mediated RNA binding protein quaking-5 on the proliferation and apoptosis of lung adenocarcinoma cell line H1299[J]. Acta Anatomica Sinica, 2020, 51(1): 40-46.

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