The Screening And Verification Of Gastric Cardiac Adenocarcinoma MicroRNA Expression Profiling

Objective: To screening the gastric cardiac adenocarcinoma microRNA expressionprofiling, provides a theoretical basis for the pathogenesis of gastric cardiacadenocarcinoma, early warning, new treatments and the development of targeted drug.Methods: From December2011, we have established gastric cardiac adenocarcinomaspecimen datebase，including tumor tissue and adjacent normal tissues, fromDecember2011. In order to ensure the quality of tissue samples, all the tumor tissueand spare adjacent normal tissues was obtained in the operating room with the30minutes from them cut from body by surgeon. Tissue samples were first placed inliquid nitrogen and transported to the laboratory, and then be placed in the-80℃refrigerator for long-term preservation.In the the first step,20patients cardia adenocarcinoma tumor tissue and correspondingadjacent normal tissues randomly selected from in the gastric cardia cancer tissue bankwere sent to KangChen Bio-tech (Shanghai, China) for screening the differencesmicroRNA expression between the gastric cardiac adenocarcinoma specimen datebaseand adjacent normal tissues though16.0version LNATMmicroRNA chip (covering1000known microRNAs).In the second step, according screening results of microRNA chip, ABI7500real-timePCR was applid to verify the differential expression of50kinds of microRNA betweenthe gastric cardiac adenocarcinoma tumor tissue and adjacent normal tissues in30patients (20patients plus another10patient randomly selected from in the gastriccardia cancer tissue bank).The screening results of microRNA chip was performed by KangChen Bio-tech(Shanghai, China). The validation results of qRT-PCR was described by mean±standard deviation format, and paired t test and Wilcoxon signed rank test was appliedto analysis the differential expression of microRNA between the gastric cardiacadenocarcinoma tumor tissue and adjacent normal tissues by SPSS statistical software. The level of statistical significance was set at α=0.05.Results: In the the first step, the screening results of microRNA chip shown that209kinds of microRNA expression differences between the gastric cardiacadenocarcinoma tumor tissue and spare adjacent normal tissues,50kinds ofmicroRNA of which were significantly higher.In the second step, by expanded the sample size,16kinds of microRNA, includingmicroRNA-20b, microRNA-195, microRNA-337, microRNA-338, microRNA-433,microRNA-10a and microRNA-645, were proved by qRT–PCR that the differentialexpression of microRNA between the gastric cardiac adenocarcinoma tumor tissue andadjacent normal tissues is still statistically significant (P <0.05), while the remaining31kinds of microRNA expression differences between the two tissues was notstatistically significant (P>0.05).Conclusions: The gastric cardiac adenocarcinoma microRNA expression profilingincluding16microRNA： microRNA-20b、 microRNA-195、 microRNA-337、microRNA-338、 microRNA-433、 microRNA-10a、 microRNA-645、 microRNA-345-5p、 microRNA-45、 microRNA-256、 microRNA-72、 microRNA-64、microRNA-27a、microRNA-100、microRNA-109and microRNA107.