| OBJECTIVEDetected the expression of Fas-associated factor1(FAF1) in differentdegrees of differentiation of Gastric Cancer Cells, selected the gastric cancercell line which the FAF1express lowest in，integrateed FAF1into the GastricCancer Cell through lentivial carrier, observed the influences of the over-expression of Fas-associated factor1(FAF1) on cell proliferation and apoptosisin human gastric cancer cell line, to explored the expression of FAF1in differentGastric Cancer Cells,and the relationship between the expression of FAF1anddegrees of differentiation of Gastric Cancer Cells, and the role it played in thedevelopment of gastric cancer.METODSThe mRNA and protein relative expression of FAF1in human gastricepithelial cell (GES-1) and gastric cancer cell lines in various degrees ofdifferentiation （well differentiated: NCI-N87, Moderately differentiated：SGC-7901, Poorly differentiated: MGC-803, undifferentia-ted: HGC-27) were detected by quantitation real-time polymerase chain reaction technology(qRT-PCR) and western blot, respectively. The gastric cancer cell line(negativecontrol group) which the FAF1express lowest in was selected to be transfectedwith empty vector particles (Empty-vector transfection group) and recombinantFAF1lentiviral particles(FAF1Overexpression group) respectively. Transfectionefficiency was detected by laser scanning confocal microscope. Proteinexpression level of FAF1was detected by Western blot. Changesof cellultrastructure were detected by transmission electron microscope. Cell cycledistribution and apoptosis were observed by Flow Cytometer. Cell proliferationwas detected by MTT assay.RASULTSPartⅠ Expression of FAF1in different Gastric Cancer Cell Lines:Compared with GES-1, the mRNA relative expression of FAF1in NCI-N87,SGC-7901, MGC-803, HGC-27were both decreased（0.76±0.06,0.61±0.03,0.50±0.05,0.40±0.03VS0.97±0.09, P＜0.01）; the protein relativeexpression of FAF1in NCI-N87, SGC-7901, MGC-803， HGC-27wereboth decreased（0.56±0.02,0.49±0.04,0.37±0.05,0.23±0.04VS0.66±0.04, P＜0.01). Morever it decreased as the degrees of differentiation of Gastric CancerCells decreased.Part Ⅱ Effects of FAF1gene over-expression on proliferation andapoptosis of Gastric Cancer Cells: The efficiency of transfection was greaterthan95%according to the green fluorescent. The expression of FAF1proteinwas obviously higher in FAF1overexpression group than the two control groups.The cell membrane of the two control groups were integrated. The cell nucleuswere unchanged. The cell ultrastructure of the two control groups had nosignificant difference. In FAF1overexpression group, cell nucleus have splited into pieces, apoptotic bodies and vacuoles have formed. Overexpressed FAF1inhibited HGC-27cell growth, induced cell apoptosis, and changed thedistribution of cell cycle. Compared to negative control group and empty-vectortransfection group, cells’ doubling time in FAF1overexpression group wassignificantly extended, cell apoptosis were significantly increased(84.66%±5.92%vs4.60%±3.80%and7.32%±3.82%, both P<0.05), thepercentage of cells in G0/G1phase was significantly decreased (46.43%±2.43%vs54.93%±3.5%and54%±0.3%, both P<0.05), the percentage of cells inG2/M phase was significantly increased (29.78%±3.91%vs19.33%±3.82%and20.93%±2.46%, P<0.05).CONCLUSION1, Compared with human gastric epithelial cell line, the mRNA and theprotein relative expression of FAF1in four kinds of gastric cancer cell lineswere both decreased, and it decreased as the degrees of differentiation of GastricCancer Cells decreased. It explained that FAF1may be a tumor suppressor genein the development of gastric cancer, and it related with the the degrees ofdifferentiation of Gastric Cancer.2, FAF1genes involved in regulating the proliferation and apoptosis ofhuman gastric cancer cell line HGC-27in vitro，it could change the distributionof cell cycle, inhibit the cells’ growth, and induce the cells’ apoptosis. FAF1plays a tumor suppressor role in the development of gastric cancer. |
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