Effect of eukaryotic expression plasmid containing methylenetetrahydrofolate reductase gene on transcriptional level of tumor-related genes in human gastric cancer cell line
- VernacularTitle:亚甲基四氢叶酸还原酶对胃癌细胞基因表达的影响
- Author:
Dan-Feng SUN
;
Jing-Yuan FANG
;
Yu-Rong WENG
;
- Publication Type:Journal Article
- Keywords:
Methylenetetrahydrofolate reductase;
Gastric cancer;
Gene expression;
Methylation
- From:
Chinese Journal of Digestion
2001;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To analyze the effect of eukaryotic plasmids containing wild (sense) or anti- sense methylenetetrahydrofolate reductase (MTHFR) gene on cell viability and transcription level of tumor related genes in human gastric cancer cell line.Methods Human gastric cancer cell line MKN-45 was cultured.Recombinant plasmids containing wild MTHFR (W) or antisense MTHFR (A) gene, pCMV-W and pCMV-A,were constructed.Then pCMV-W,pCMV-A and pCMV blank plasmid were transfected into MKN45 cells respectively by using lipofect.Cell viability was analyzed by 3-(4,5-bime- thylthiazolyl-2)-2,5-diphenyhetrazolium dromide(MTT).The transcription levels of Dnmt 1,c-myc, p21~(WAF1) and hMLH1 genes were detected by real-time polymerase chain reaction(PCR).Results Cell vi- ability remarkably increased in those transfected with wild MTHFR (P＜0.01),which was contrary to those transfected with antisense MTHFR(P＜0.01).The expression of those tumor related genes mRNAs were all remarkably decreased in the MKN45-W cells in comparison with those in the MKN45-pCMV cells.No significant difference in the expressions of those tumor related genes mRNAs were found between the MKN45 cells transfected with pCMV-A and blank pCMV.Conclusion MTHFR influences cell viability and the expres- sion level of tumor related genes in human gastric cancer cell line MKN45.
