Effects Of RASSF1aA On MicroRNA Expression Profiles Of Human Gastric Carcinoma SGC-7901Cells

Objectives: The purpose of the study is to establish human gastriccarcinoma cell line SGC-7901with high expression of RASSF1A and toinvestigate its roles on microRNA expression profiles of gastric cancinomacells.Methods: pcDNA3.1-RASSF1A and pcDNA3.1were introduced intohuman gastric carcinoma SGC-7901cell line by lipofectin transfection, andthe SGC-7901cells stably expressing RASSF1A gene were established byG418selection. The expression of RASSF1A was detected by reversetranscription-polymerase chain reaction (RT-PCR) and western blotting.miRCURY LNA Array (Exiqon, Denmark) was used to identify differentiallyexpressed microRNAs between over expression RASSF1A genes of pcDNA3.1-RASSF1A-SGC-7901cells and pcDNA3.1-SGC-7901from control.Dateanalysis was performed with the Axon GenePix4000B and GenePix pro V6.0was used to read the raw intensity of the image. Finally, To confirm the resultsof miRNA Array.We chosed three of the differential expressed miRNA,detected miR-181a,miR-181b and miR-711expression by real time RT-PCRassay between pcDNA3.1-SGC-7901from control and over expressionRASSF1A genes of pcDNA3.1-RASSF1A-SGC7901cells.Results: Human gastric carcinoma SGC-7901cells stably over expressingRASSF1A gene were established by lipofection mediated transfection.Fourteen microRNA were differentially expressed in over expressingRASSF1A gene SGC-7901cells compared with vector transfected SGC-7901cells by miRCURY LNA Array.Seven were up-expressed(miRPlus-E1153,miR-1207-3p, miRPlus-E1225, miRPlus-E1133, miRPlus-E1077, miR-377,miR-711) and seven were under-expressed (miR-181a, miR-181b, miR-330-5p,miRPlus-F1086, miR-483-5p, miR-132,miRPlus-E1026)in over RASSF1Agenes of SGC-7901. miR-181a, miR-181b and miR-711which were selectedfor real-time RT-PCR verification, real-time RT-PCR data were concordantwith the microarray data. Conclusions:1.Over expression of RASSF1A tumor suppressor genes canalter microRNA expression profiles of human gastric carcinoma SGC-7901cells.2.Aberrant expression of miR-181a,miR-181b and miR-711associate withgastric carcinoma.