Construction of SHIP1 overexpressed NSCLC cell line and the effect of SHIP1 on NSCLC cell proliferation
10.3872/j.issn.1007-385x.2018.07.004
- VernacularTitle:SHIP1基因过表达非小细胞肺癌细胞系的建立及其对细胞增殖能力的 影响
- Author:
TAO Xingyu
1
,
2
;
FU Qiaofen
3
;
WANG Ying
3
;
SONG Xin
1
,
2
Author Information
1. 1. Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong, China
2. 2. Department of Cancer Biotherapy Center, the Third Affiliated Hospital of Kunming Medical University and Tumor Hospital ofYunnan Province, Kunming 650118,Yunnan, China
3. Department of Cancer Biotherapy Center, the Third Affiliated Hospital of Kunming Medical University and Tumor Hospital ofYunnan Province, Kunming 650118,Yunnan, China
- Publication Type:Journal Article
- Keywords:
SHIP1;
non-small cell lung cancer;
gene recombination;
stable overexpression;
cell proliferation
- From:
Chinese Journal of Cancer Biotherapy
2018;25(7):680-686
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To observe the effect of SHIP1 on NSCLC cell proliferation. Methods: The CDS region of human SHIP1 gene was obtained by inquiring NCBI Gene database and was inserted into the vector pTSB-CMV-MCS-SBP-3Flag-EGFP to construct SHIP1 over-expression plasmid, which was further used to construct SHIP1 overexpression lentivirus. SHIP1 over-expressed lentiviruses were used to transfect A549, SPCA-1 and PC-9 cell lines to construct SHIP1 overexpressed NSCLC cell line. Western blotting and qRT-PCR were used to determine the protein and mRNAexpression of SHIP1. The MTT assay and Clone formation assay were used to examine the cell proliferation ability and clone formation ability of PC-9 cells overexpressed SHIP1; Western blotting was performed to examine the level of AP-1 proteins. Results: The sequencing result suggested that the SHIP1 eukaryotic over-expression plasmid was successfully constructed. A519, SPCA-1 and PC-9 cells with SHIP1 over-expression were observed to display uniform green fluorescence under fluorescent microscopy. Compared with negative control group, the mRNA and protein levels of SHIP1 were significantly increased in SHIP1 overexpressed cells (all P<0.01). The over-expression of SHIP1 suppressed the abilities of proliferation and clone formation in PC-9 cells (all P<0.01), and down-regulated the expression of p-c-Jun and FosB etc. Conclusion: The SHIP1 overexpressed NSCLC cell lines were successfully established, and the over-expression of SHIP1 suppressed the cell proliferation ability by inhibitingAP-1 proteins in NSCLC cell lines.
- Full text:20180704.pdf
