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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
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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.
作者 苏锐 李英健 朱志达 赵恩宏 Su Rui;Li Yingjian;Zhu Zhida;Zhao Enhong(Department of Gastrointestinal Surgery,Affiliated Hospital of Chengde Medical College,Chengde 067000,China)
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)。
关键词 胃肿瘤 细胞增殖 细胞凋亡 miR-496 Stomach neoplasms Cell proliferation Apoptosis miR-496
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