Construction and transfection of the recombinant plasmid pEGFP-C1-antisense survivin / 中华消化杂志

ABSTRACT

Objective To construct recombinant plasmid pEGFP C1 antisense survivin and than transfect it to MKN 45 gastric cancer cells for studying the expression of survivin mRNA and its effects on apoptosis of cancer cells. Methods Recombinant pEGFP C1 antisense survivin plasmid was cloned and transfected to MKN 45 gastric cancer cells by lipofectamine. Apoptosis of gastric cancer cells was assayed by flow cytometry. The levels of survivin mRNA before and after transfection were determined by reverse transcription polymerase chain reaction analysis. Results After recombinant pEGFP C1 antisense survivin plasmid was transfected, the apoptosis of MKN 45 cells was increase, the cell number in G 2/M phase was decreased, and the level of survivin mRNA was inhibited. Conclusions pEGFP C1 antisense suvivin can promote the apoptosis of MKN 45 gastric cancer cells, that might attribute to the inhibition on cell proliferation and expression of survivin mRNA.