Construction of Caveolin-1 RNA interfering eukaryotic expressing vector and its influence on biological behavior of human gastric cancer cells / 肿瘤研究与临床

ABSTRACT

Objective To design siRNA for Caveolin-1(CAV1) and construct eukaryotic expression vector Dsilence4.1-CMV-neo-CAV1, which is transfected into gastric cancer cell line SGC7901 to study the effects of inhibition of CAV1 expression with siRNA on biological behavior of gastric cell line SGC7901.Methods Design and chemosynthesize three pairs of double strands siRNA based on the neucleotide sequence of CAV1 gene provided by GeneBank.The siRNAs were transient transfected into SGC7901.The CAV1 expression of transfected cells were detected respectively by semi-quantity RT-PCR to sieve a valid siRNA sequence.Design and synthesized DNA sequence which can express small hairpin RNA(shRNA),construct RNA interfering eukaryotic expression vector of CAV1 gene.The SGC7901 Was stably transfected and was detected the expression of CAV1 gene by RT-PCR.The influence on the gastric cell growth and pmliferation after CAV1 was inhibited Was detected by cell growth curve, clone formation test and FCM.Results The CAV1 RNA interfering eukaryotic expression vector psilence4.1-CMV-neo-CAV1 was successfully constructed,which can significantly reduce the expression of CAV1 after stably transfected to target cells,and in contrast to Dsilence4.1-CMV-neo transfection group,PI and colony for mution rate were increased(t=7.98,P<0.05;t=13.19,P<0.01),the growth speed Was up.Conclusion The growth and proliferation ability of tumor is remarkablv accelerated by inhibiting the expression of CAV1 of gastric cell by RNAi.