Influence Of Combined Folate Deficiency With PLK1 SiRNA In The Polymorphism Of Gastric Cancer Cells

Objective: To observe the PLK1 siRNA, folate deficiency and their combination to the expression of FR-?,Bcl-2,P53,MTHFR polymorphism of gastric cancer cells SGC-7901,BGC-823,N87 and gastric mucosal cells GES-1.And whether these effects disappear after folate deficiency environmental is corrected.Methods: The gastric cancer cell SGC-7901, BGC-823, N87 and gastric mucosal cell GES-1 were placed in PLK1 siRNA interference, folic acid deficiency, PLK1 siRNA interference joint folate deficiency and after folic acid deficiency environmental restoration then PLK1 siRNA interference conditions cultured for a period of time, First of all,detected each cell line FR-?, Bcl-2, P53 m RNA expression by Real-time PCR. Then detected each cell line FR-?, Bcl-2, P53 protein expression by Western blot. Finally, experimental cell using PCR-RFLP method and gene chip technology detected the MTHFR gene polymorphism.Results: 1.through Real-time PCR detection, after PLK1 siRNA interference, experimental cell FR-? and Bcl-2 m RNA expression was significantly lower than the control group, the P53 m RNA expression was significantly higher than the control group(p<0.05); When folic acid deficiency, experimental cell FR-? m RNA expression was significantly higher than the control group(p <0.05), however, P53 and Bcl-2 m RNA expression was no significant change(p> 0.05). When combined with a lack of folic acid and PLK1 siRNA interference, both exhibiting antagonism and tend to PLK1 siRNA interference separately;But the antagonism between the two disappear after folic acid deficiency environmental restoration then PLK1 siRNA interference conditions.2. By Western blot detection, after PLK1 siRNA interference, experimental cell FR-? and Bcl-2 protein expression was significantly lower than the control group, the P53 protein expression was significantly higher than the control group(p <0.05). When folic acid deficiency, the results contrast. When combined with a lack of folic acid and PLK1 siRNA interference, both exhibiting antagonism and tend to PLK1 siRNA interference separately;But the antagonism between the two disappear after folic acid deficiency environmental restoration then PLK1 siRNA interference conditions.3. MTHFR gene was successfully amplified two fragments containing 667 points and containing 1298 points, through gene sequencing, In addition to the 667-point cells NCI-N87 is no mutation,the rest cells all by C to T mutation, while in the 1298 point, none of the experimental cell gene mutation.Conclusions: 1, PLK1 siRNA and folate deficiency may influence the development of gastric cancer by Bcl-2, P53 and FR-? gene.2.PLK1 siRNA interfere with folate deficiency when combined antagonism, but the impact of short-term folic acid deficiency is reversible, folic acid supplements may prevent cancerous transformation of normal gastric mucosa cells, inhibit progression of gastric cancer cells;3. FR-? is a potential tumor markers, may similar with the oncogenes Bcl-2,opposite to tumor suppressor gene P53,is a potential oncogenes; 4. MTHFR variant has many, MTHFR C677 T points polymorphism may be a genetic risk factor of gastric cancer.