论文部分内容阅读
目的:观察环状RNA(circRNA) cRAPGEF5对胃癌细胞增殖、侵袭和转移的影响。方法:选取新乡医学院第一附属医院2018年1月至2020年1月手术切除的143例胃癌组织和癌旁组织作为研究对象，采用转录组织化学测序和荧光定量聚合酶链反应(FQ-PCR)分析cRAPGEF5表达水平；根据过表达的circRNA的类型分为对照组和实验组，采用慢病毒在胃癌细胞系BGC-823建立circRNA对照和cRAPGEF5过表达细胞系(对照组和实验组)。采用细胞计数试剂盒(CCK-8)和克隆形成实验分析两组细胞增殖能力；采用Transwell分析细胞侵袭能力，体内移植瘤实验分析肿瘤生长和转移能力。组间数据比较采用n t检验。n 结果:转录组学显示胃癌组织和癌旁组织cRAPGEF5存在显著差异表达。胃癌组织中cRAPGEF5表达水平(0.39±0.11)低于癌旁组织中cRAPGEF5表达水平(1.19±0.19)，差异有统计学意义(n t＝3.683，n P<0.05)。本研究成功采用慢病毒在胃癌细胞系BGC-823建立cRAPGEF5过表达稳定细胞系。实验组细胞吸光度(n A)值(1.21±0.16)低于对照组细胞n A值(1.85±0.21)，差异有统计学意义(n t＝2.901，n P<0.05)。克隆形成实验结果显示，实验组细胞克隆形成率[(25.19±2.09)%]低于对照组细胞克隆形成率[(51.32±4.89)%]，差异有统计学意义(n t＝4.129，n P<0.05)。实验组细胞侵袭数量[(43.09±5.48)个]低于对照组细胞侵袭数量[(98.39±8.99)个]，差异有统计学意义(n t＝5.099，n P<0.05)。实验组细胞接种裸鼠30 d后肿瘤体积和肿瘤重量[(1 029.89±159.33) mmn 3；(1.69±0.34) g]低于对照组肿瘤体积和肿瘤重量[(698.74±76.82) mmn 3；(0.89±0.26) g]，差异有统计学意义(n t＝5.091，n P<0.05；n t＝3.105，n P<0.05)。对照组细胞接种裸鼠30 d后腋下淋巴结转移率为65.00%(13/20)，明显高于实验组腋下淋巴结转移率20.00%(4/20)，差异有统计学意义(n χ2＝2.980，n P<0.05)。n 结论:cRAPGEF5在胃癌中呈低表达，参与胃癌的增殖、侵袭和转移。“,”Objective:To investigate the effect of circular RNA (circRNA) cRAPGEF5 on the proliferation, invasion and metastasis of gastric cancer cells.Methods:A total of 143 cases of gastric cancer and paracancerous tissues from January 2018 to January 2020 in our hospital were selected as the research objects. The expression level of cRAPGEF5 was analyzed by transcriptomics and fluorescence quantitative polymerase chain reaction (PCR). CircRNA control and circRNA cRAPGEF5 overexpression cell lines (control group and experimental group) were established by lentivirus in gastric cancer cell line BGC-823. The proliferation ability was analyzed by cell counting kit-8 (CCK-8) assay and clone formation assay was done in the two groups by plate clone formation experiment. The invasion ability was analyzed by Transwell. The tumor growth and metastasis ability were analyzed by n in vivo transplantation tumor test.n T test was used to compare the data between groups, and the difference was statistically significant whenn P<0.05.n Results:Transcriptomics results showed that the expression of cRAPGEF5 was significantly different between gastric cancer and adjacent tissues. As compared with the adjacent tissues (1.19±0.19), the expression level of cRAPGEF5 in gastric cancer tissues (0.39±0.11) was significantly decreased (n t=3.683, n P<0.05). In this study, we successfully established a stable cell line with overexpression of cRAPGEF5 in gastric cancer cell line BGC-823. As compared with the control group (1.85±0.21), the cell absorbance valuen A (1.21±0.16) in the experimental group significantly decreased (n t=2.901, n P<0.05). As compared with the control group [(51.32±4.89)%], the clone formation rate of the experimental group [(25.19±2.09)%] was significantly decreased (n t=4.129, n P<0.05). As compared with the control group (98.39±8.99), the number of invading cells in the experimental group (43.09±5.48) significantly decreased (n t=5.099, n P<0.05). The tumor volume and tumor weight in the experimental group [(1 029.89±159.33) mmn 3, (1.69±0.34) g] were significantly reduced as compared with those in the control group [(698.74±76.82) mmn 3, (0.89±0.26) g, n t=5.091, n P<0.05;n t=3.105, n P<0.05]. The axillary lymph node metastasis rate was 65.00% (13/20) in the control group, significantly higher than 20.00% (4/20) in the experimental group after 30 days of cell inoculation (n χ2=2.980, n P<0.05).n Conclusion:The expression of cRAPGEF5 is low in gastric cancer, which is involved in the proliferation, invasion and metastasis of gastric cancer.