Preliminary Study On FGGY Function In Gastric Cancer

Background It is reported that 2012 global cancer about 14.1 million cases and 8.2 million deaths,of which the incidence of gastric cancer ranked fifth in all cancer,the mortality rate is third.At present the international adoption of gastric cancer treatment program for the operation of the main comprehensive treatment.Some scholars believe that advanced gastric cancer stressed the scope of the increase in cleaning,combined with organ resection and other surgical methods can not bring more benefits to patients.And for patients with advanced surgical treatment and radiotherapy and chemotherapy methods such as poor efficacy,5-year survival rate of patients is still less than 30%.It is urgent to seek new and effective treatment of gastric cancer,including tumor gene therapy.Inactivation of tumor suppressor gene and tumor growth are closely related to the normal tumor suppressor gene into tumor cells to compensate for and replace the mutant or missing tumor suppressor gene,to inhibit tumor growth or reverse malignant tumor tumor suppressor gene therapy strategy,Become an important therapeutic mode of tumor gene therapy.In this paper,by means of lentivirus infection,Western blot,soft agar clone formation test and other methods to detect whether FGGY is a candidate gastric cancer specific tumor suppressor gene.? The detection of different cell lines FGGY protein expression differencesMethods1.Western blot detection of normal gastric mucosa cells and other sources of normal cell lines FGGY protein expression differences2.Western blot detection of normal gastric mucosa cells and gastric cancer cell line FGGY protein expression differencesRes?lts1.Western blot detection of normal gastric mucosa cells and other normal cell lines FGGY protein expression differences Western blot showed that FGGY protein was highly expressed in GES-1,JB6,Ha Ca T,HIEC and CCD-18 Co cell lines.2.Western blot detection of normal gastric mucosa cells and gastric cancer cell line FGGY protein expression differences Western blot showed that SGC7901,BGC803,MGC823 and SGC7901 showed low expression or no expression.? The effect of FGGY expression on the growth of gastric cancer cells.Methods1.The FGGY plasmid into p CMV to construct the overexpression vector,sent to Shanghai biopsy validation.2.We will get FGGY low expression or not express gastric cancer cell lines,transfected with p CMV-FGGY,transfected with p CMV-Mock as control group,WB to determine FGGY protein overexpression efficiency.The effect of FGGY on non-anchoring growth of gastric cancer cells was detected by soft agar clone formation assay.Each gastric cancer cell was divided into two groups: pCMV-Mock group,pCMV-FGGY group,c?ltured in cell incubator for7-14 days,and the number of clones was observed under microscope.Res?lts1.Gastric cancer cells transfected with p CMV-FGGY,FGGY protein expression was significantly increased.2.Found that expression of FGGY significantly inhibited the non-anchoring growth of cancer cells,clonal formation significantly reduced.? Effects of gene silencing FGGY on normal cell transformation induced by EGFMethods1.Design FGGY sh RNA primer,do PCR to get fragments,cloned to plko expression vector to construct FGGY sh RNA.The expression of FGGY protein in WGG gene was detected by silencing stable cell line and FGGY.The expression of FGGY protein in WGG gene was detected by the expression of FGG and sh FGGY infected with JB6 and puromycin.2.Soft agar clone formation assay to detect gene silencing FGGY on EGF-induced cell transformation.Res?lts1.After gene silencing,the expression of FGGY protein in JB6 cells was significantly decreased.2.Found that down-reg?lation of FGGY significantly enhanced EGF-induced cell transformation,clonal formation time in advance,the number significantly increased the diameter significantly increased.Conclusion1.Cell experiments confirmed that FGGY gene inhibition of gastric cancer cell proliferation,and its invasion,migration ability to inhibit.2.FGGY gene is a potential tumor suppressor gene,in the occurrence and development of gastric cancer reg?lation inhibitory effect,may become a potential target of gastric cancer gene therapy.