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Archive > Volume 25 Issue 7 > 2018,25(7):680-686. DOI:10.3872/j.issn.1007-385X.2018.07.004 Prev Next
Basic Research
Construction of SHIP1 overexpressed NSCLC cell line and the effect of SHIP1 on NSCLC cell proliferation
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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 mRNA expression 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 inhibiting AP-1 proteins in NSCLC cell lines.
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Project Supported:
Project supported by the National Natural Science Foundation of China (U1502222, No.81702295), the Project of Health Science and Technology of Yunnan Province (No.2017NS177, No.2016NS113), the Project of Science and Technology of Yunnan Province (No.2014FB064), and the China Postdoctoral Science Foundation (No.2017M613008)
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