miR-496通过调控 LYN对胃癌细胞增殖和凋亡的影响  

Effects of miR-496 on proliferation and apoptosis of gastric cancer cells through regulation of LYN

作　　者：苏锐[1] 李英健[1] 朱志达 赵恩宏[1] Su Rui;Li Yingjian;Zhu Zhida;Zhao Enhong(Department of Gastrointestinal Surgery,Affiliated Hospital of Chengde Medical College,Chengde 067000,China)

机构地区：[1]承德医学院附属医院胃肠外科,承德067000

出　　处：《国际外科学杂志》2023年第9期591-595,F0003,F0004,共7页International Journal of Surgery

基　　金：承德市科技计划项目(202109A186)。

摘　　要：目的研究miR-496在胃癌细胞中的表达情况, 并探讨其在胃癌细胞增殖和凋亡过程中的作用及机制。方法实时荧光定量PCR(qPCR)检测miR-496在正常胃上皮细胞株和胃癌细胞系AGS、MKN45中的的表达情况。在表达水平最低的AGS细胞中敲低miR-496, 同时设置阴性对照组和空白对照组。分别用CCK8实验和流式细胞仪检测细胞增殖和凋亡。使用生物信息学软件筛选出miR-496的靶基因LYN, QPCR检测转染miR-496对LYN表达的影响, 随后进行了Rescure拯救实验, 进一步研究miR-496通过调控LYN对胃癌细胞的作用机制。利用GraphPad Prism 9软件进行数据分析, 计量资料以均数±标准差(±s)表示, 组间比较采用t检验。结果 miR-496在AGS、MKN45中表达显著低于胃正常上皮细胞(P<0.05)。通过转染过表达miR-496后, 可以抑制AGS细胞的增殖, 同时可以使AGS细胞的凋亡比例明显升高(P<0.05)。qPCR结果显示, miR-496过表达组能够抑制LYN的表达(P<0.05)。生物信息学分析显示, miR-496与LYN激酶(LYN) 3’UTR区域结合, 通过过表达miR-496可抑制AGS细胞中LYN的表达, 而CCK8拯救实验显示过表达LYN能够解除miR-496对细胞增殖的抑制作用;流式凋亡检测显示表达LYN能够解除miR-496对细胞凋亡的促进作用(P<0.05)。结论 miR-496在胃癌细胞中低表达, 其通过靶向胃癌细胞中LYN的表达抑制胃癌细胞的增殖, 促进胃癌细胞的凋亡。Objective To study the expression of miR-496 in gastric cancer cells,and explore its role and mechanism in the proliferation and apoptosis of gastric cancer cells.Methods Real-time fluorescence quantitative PCR(qPCR)was used to detect the expression of miR-496 in normal gastric epithelial cell lines and gastric cancer cell lines AGS and MKN45.miR-496 was knocked down in AGS cells with the lowest expression level,and a negative control group and a blank control group were set up.Cell proliferation and apoptosis were detected by CCK8 assay and flow cytometry.LYN,the target gene of miR-496,was screened using bioinformatics software,and the effect of transfection of miR-496 on LYN expression was detected by qPCR.Subsequently,rescure experiment was conducted to further study the mechanism of miR-496 on gastric cancer cells through regulation of LYN.Data were analyzed by GraphPad Prism 9 software.Measurement data were presented as mean±standard deviation,and the comparison between the two groups was performed by t test.Results The expression of miR-496 in AGS and MKN45 was significantly lower than that in normal gastric epithelial cells(P<0.05).After overexpression of miR-496,the proliferation of AGS cells could be inhibited and the apoptosis ratio of AGS cells could be significantly increased(P<0.05).QPCR results showed that miR-496 overexpression group could inhibit the expression of LYN(P<0.05).Bioinformatics analysis showed that miR-496 binds to LYN kinase(LYN)3′UTR region,and overexpression of miR-496 can inhibit the expression of LYN in AGS cells,while CCK8 rescue experiment showed that overexpression of LYN could remove the inhibitory effect of miR-496 on cell proliferation.Flow cytometry showed that LYN expression could cancel the promoting effect of miR-496 on apoptosis(P<0.05).Conclusion miR-496 is low expressed in gastric cancer cells,and it inhibits the proliferation and promotes apoptosis of gastric cancer cells by targeting the expression of LYN in gastric cancer cells.

关 键 词：胃肿瘤 细胞增殖 细胞凋亡 miR-496 

分 类 号：R735.2[医药卫生—肿瘤]