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Effect of STGC3 Gene Deletion Mutant on the Growth of CNE2 Cell Line

Author: LiLi
Tutor: HeXiuSheng
School: Nanhua University
Course: Pathology and Pathophysiology
Keywords: nasopharyngeal carcinoma CNE2 cell line STGC3 gene gene deletion mutant LG domain
CLC: R739.63
Type: Master's thesis
Year: 2011
Downloads: 4
Quote: 0
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Abstract


Objective: The Present study was to investigate functional domain of STGC3 gene, through observing the effect of Laminin G domain deletion mutant in cell growth and proliferation.Methods: A recombinant plasmid encoding the STGC3 deletion mutant (pcDNA3.1（+）-STGC3 △142AA) was verified by double restriction digestion and DNA sequencing, and then introduced into CNE2 cells by lipofectin transfection. Positive clones were selected in the present of G418, the expression of the deletion mutant was detected by reverse transcription polymerase chain reaction （RT-PCR）. Cell proliferation changes were measured by drawing growth curve and experimenting plate clone formation. The cell cycle distribution was tested by flow cytometry.Results: The recombinant plasmid pcDNA3.1（+）-STGC3△142AA was verified to contain STGC3 △142AA by double digestion and DNA sequencing. The stable CNE2 cell lines contained these recombinants were established and verified by RT-PCR. This cell line showed enhanced proliferation and anchorage-independent growth as revealed by MTT assay and plate clone formation assay, compared with the wild type control. FCM analysis indicated that G0/G1 Phrase cell rate of CNE2/STGC3△142AA was lower markedly than that of CNE2/STGC3.Conclusion:1. Recombinant expression vector pcDNA3.1（+）-STGC3△142AA was successfully constructed.2. Deleting LG domain of STGC3 gene leads to decrease in negative regulation of the gene on growth and proliferation ability of CNE2 cell line. 3. LG domain was the key region of STGC3 proteinic.

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