Study Of The Level Of NF-κB Expression In The Gastric Cancer Cell Lines Infected By Hp From Patients Of Coalmine Workers

Objective To study the level of NF-κB expression in gastric cancer cell lines infectied by Hp from coal mine workers and general population , explore the relationship between different Hp strains and NF-κB,and the activation rules of NF -κB in gastric cancer cell lines with different differentiation, to provide clues for mechanism study of Hp promoting gastric cancer and scientific basis for prevention and control of gastric cancer .Methods Strains groups: collect gastric mucosal epithelium cells of coal miners and the general population with Hp related stomach diseases for Hp cultivation, According to CagA gene, Hp was divided into four groups: coal miner HpCagA~ + strains, coal miner HpCagA~ -strains, general population HpCagA~ + strains, general population HpCagA~- strains.Add NCTC11637 standard strains,total five groups.Cell groups: Human undifferentiated, low differentiated, High differentiated gastric cancer cell lines(BGC-823,MKN-45,MKN-28),total three groups. With 100:1 ratio,five Hp strains were added to BGC-823,MKN-45,MKN-28 cell lines parallelly, one bottle cells per strains. Additionally,one bottle blank control cells per cell lines.Total eighteen-bottle cells.Repeat above experiment three times.After Hp strains infected gastric cancer cell lines,cultivate them 24 hours in carbon dioxide incubator under the condition of 37℃.Then crack cells, detect protein content with BCA method. Test the expression level of NF -κB with Western Blot. Takingβ-actin as internal reference, to compare the differences of NF -κB expression level.Results 1,Successful cultured four kinds of Hp strains: HpCagA~ + strains, CagA~–strains from coal miner workers and general population with Hp related stomach diseases and human three kinds undifferentiated, low differentiated, High differentiated gastric cancer cell lines(BGC-823,MKN-45,MKN-28). 2,Without Hp infection ,the content differences of NF-κB in BGC-823,MKN-45,MKN-28 cell lines have no statistical significance.3,After Hp infection, the content differences of NF-κB in BGC-823,MKN-45,MKN-28 cell lines have no statistical significance also.4,The expression content of NF-κB in cell lines infected by Hp is higher than that of NF-κB in cell lines with no Hp infection.The differences has statistical significance.5,The expression content of NF-κB in cells infected by HpCagA+strains,HpCagA- strains is higher than that of NF-κB in cells of blank group, P<0.05. The expression content differences of NF-κB between HpCagA+strains group and HpCagA- strains group have no statistical significance.6,The expression content of NF-κB between blank group and general population HpCagA- strains group are lower than that of NF-κB of NCTC11637 standard strains group,coal mine worker HpCagA+ strains group,general population HpCagA+strains group, coal mine worker HpCagA- strains group.The differences between the front two groups have no statistical significance.The differences among the back four groups have no statistical significance.7,The expression contents of NF-κB of NCTC11637 standard strains group,coal mine worker Hp strains group,general population Hp strains group are higher than that of NF-κB of blank group, the differences have statistical significance. The differences among the front three groups have no statistical significance.Conclusion The rise of NF-κB expression level among three gastric cancer cell lines of BGC-823,MKN-45,MKN-28 is related to Hp infectin,not associated with gastric cancer cell differentiation degree , CagA and the resource of Hp.