The Study Of The Role Of MiR-1248 And MiR-193b-3p On Apoptosis Processes Induced By Tetrandrine In Human Gastric Cancer

Objective To investigate the growth inhibitory and apoptosis effects of the Low expression of mi R-1248 and over expression of mi R-193b-3p on the gastric cancer cells(SGC-7901,HGC-27)by Tet.Methods The proliferation of gastric cancer cells treated with Tet was detected by WST-1,and the cell cycle progression and cell apoptosis were determined by flow cytometry.The use of high-throughput sequencing technology,mi RNA-Seq and RNA-Seq were analyzed to detect the different mi RNAs and genes after treatment with Tet.And the expression of mi R-1248 and mi R-193b-3p in gastric cancer cells were confirmed by Real-time RT-PCR.The Low expression of mi R-1248 and over expression of mi R-193b-3p were transfected in gastric cancer cells,respectively,to explore the proliferation and cell apoptosis of gastric cancer cells by WST-1 and flow cytometry.And cells migration and invasion abilities were detected by wound-healing assay,Transwell migration assay and Transwell invasion assay.Results一 、 The different concentrations of Tet can inhibit gastric cancer cells(SGC-7901,HGC-27)proliferation in concentration dependent growth.When the concentration of 10.00μmol/L,the inhibition rate of SGC-7901 cells was68.32%,when the concentration of 2.50μmol/L,the inhibition rate of HGC-27 cells was 60.75%.二、Compared with solvent group,The cell cycle distribution obtained by flow cytometry showed that Tet arrested the cell of gastric cancer cells at S phase after 48 hours,and the proportion of S phase increased by 9.05%(P<0.01)and 16.73%(P<0.01)respectively.三、The apoptosis rate of gastric cancer SGC-7901 andr HGC-27 cells increased by 5.37%(P<0.01)and 11.05%(P<0.01),compared with the control group.It is suggested that Tet can induce gastric cancer cell apoptosis.四、The result of High throughput sequencing technologies showed that the mi RNA expression of gastric cancer cells was significantly different after treatment Tet compared with the control group,which 93 up-regulated genes and 139down-regulated genes were identified.According to the co-expression analysis of varied mi RNAs and RNAs,totally 81 mi RNA-RNA pairs were identified.The results of Real-time RT-PCR showed that the expression ratio of mi R-1248 after treatment with Tet was 9.9 × 10-3-fold(Tet/control group),while the expression ratio of mi R-193b-3p was about 103-fold(Tet / control group).五、Compared with inhibitor NC,the cell viability of SGC-7901 and HGC-27 cells were significantly decreased by 40.15% and 20.81%,respectively after 72 h transfection with mi R-1248 inhibitor by WST-1.Compared with mimic NC,the number of SGC-7901 and HGC-27 cells decreased by 33.03% and 22.92%,respectively after transfection with mi R-193b-3p mimic.六 、 The cell cycle distribution obtained by flow cytometry showed that mi R-1248 inhibitor and mi R-193b-3p mimic promoted the apoptosis of gastric cancer cells after 72 h,respectively.The apoptosis rate of mi R-1248 inhibitor was 18.03%(P<0.01)and 13.13%(P<0.05).The apoptosis rates of mi R-193b-3p mimic was 36.1%(P<0.01)and 36.1%(P<0.01).七、After transfection with mi R-1248 inhibitor and mi R-193b-3p mimic for 24 h and 48 h,the metastasis ability of gastric cancer cells were declined.The cell healing rate of SGC-7901 cells was reduced by 2.93% and 23.28% at 24 h and 48 h after transfection of mi R-1248 inhibitor,respectively.The cell healing rate of HGC-27 cells decreased by 32.51%(P<0.05)and 28.48%(P<0.05).After transfected with mi R-193b-3p mimic,the cell healing rate of SGC-7901 cells was decreased by18.88%(P<0.05)and 21.06%(P<0.05)at 24 h and 48 h.HGC-27 was reduced by25.47%(P <0.05)and 25.97%(P <0.05),respectively.八、The Transwell migration assay show that transfection of mi R-1248 inhibitor and mi R-193b-3p mimic,compared with the control group,the average cell number of SGC-7901 and HGC-27 cells were significantly reduced.After transfection of mi R-1248 inhibitor,the percentage of SGC-7901 and HGC-27 cells were decreased by 67.3%(P<0.01)and 83.0%(P<0.01).After transfection with mi R-193b-3p mimic,the percentage of SGC-7901 and HGC-27 cells were decreased by 54.3%(P<0.05)and 70.4%(P<0.01),respectively.九、The invasion ability of gastric cancer cells was inhibited after transfection with mi R-1248 inhibitor and mi R-193b-3p mimic by Transwell invasion assay.Compared with inhibitor NC group,the invasion inhibition rates of SGC-7901 and HGC-27 cells were 40.24% and 69.92% after transfected mi R-1248 inhibitor(P<0.01).After transfected with mi R-193b-3p mimic,SGC-7901 and HGC-27 cells were the invasion inhibition rate of 64.30%(P<0.01)and 60.84%(P<0.05)respectively.Conclusions一、Tet can effectively inhibit the proliferation of gastric cancer cells and promote apoptosis.二、The gastric cancer cells were promoted apoptosis by transfected with mi R-1248 inhibitor and mi R-193b-3p mimic.mi R-1248 inhibitor and mi R-193b-3p mimic has the ability to inhibit the migration and invasion of gastric cancer cells..三、Tet may inhibit the proliferation of gastric cancer cells by regulating the expression of mi R-1248 and mi R-193b-3p.