环状RNA circFOXO3在胃癌组织中的表达及其对胃癌细胞增殖和侵袭转移能力的影响

Expression of circular RNA circFOXO3 in gastric cancer tissues and its effects on proliferation, migration and invasion of gastric cancer cells

目的观察circFOXO3在胃癌组织的表达及对胃癌细胞增殖和侵袭能力的影响。方法收集2017年8月至2018年10月于河南省人民医院胃肠外科进行胃癌根治术的48例胃癌患者组织标本,通过实时定量反转录聚合酶链反应(RT-qPCR)检测circFOXO3在胃癌组织中的表达,分析circFOXO3表达与胃癌组织临床病理特征之间的相关性。构建circFOXO3过表达和对照质粒(pEX-circFOXO3、pEX-NC)、敲减和对照载体(si-circFOXO3、si-NC),分别设为过表达组和对照组、敲减组和对照组,采用细胞计数(CCK-8)、Transwell侵袭和转移实验检测胃癌细胞的增殖、侵袭和转移能力。计量资料组间比较采用t检验,计数资料采用χ2检验。结果circFOXO3在胃癌组织中高表达率(29/48,60.42%)显著高于癌旁正常组织(16/48,33.33%)(χ2=7.069,P<0.05),且其表达与组织分化(χ2=5.976,P<0.05)、浸润深度(χ2=4.409,P<0.05)、TNM分期(χ2=5.581,P<0.05)呈正相关。CCK-8、Transwell侵袭和转移实验结果表明,过表达circFOXO3后AGS细胞增殖(24 h:0.95±0.08;48 h:1.76±0.16)、侵袭[(157±11)个]和迁移能力[(186±14)个]均较对照组[24 h:0.58±0.04,t=7.165,P<0.05;48 h:0.88±0.07,t=8.728,P<0.05;侵袭为(85±7)个,t=9.565,P<0.05;迁移为(94±8)个,t=9.882,P<0.05]显著提高;敲减circFOXO3后SGC-7901细胞增殖(24 h:0.46±0.02,48 h:0.73±0.05)、侵袭[(53±4)个]和迁移[(65±5)个]能力较对照组[24 h:0.63±0.04,t=6.584,P<0.05;48 h:1.34±0.14,t=7.107,P<0.05;侵袭为(96±8)个,t=8.327,P<0.051;迁移为(114±9)个,t=8.243,P<0.05]均明显降低,差异有统计学意义。结论circFOXO3在胃癌组织中显著高表达,且可促进胃癌细胞增殖和侵袭转移。

Objective To observe the expression of circFOXO3 in gastric cancer tissues and its effects on the proliferation,migration and invasion of gastric cancer cells.Methods 48 gastric cancer samples were collected from August 2017 to October 2018 in our hospital.The expression of circFOXO3 in gastric cancer tissues was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).The correlations between the expression of circFOXO3 and the clinicopathological characteristics of gastric cancer were analyzed.The overexpression and control plasmids(pEX-circFOXO3,pEX-NC),knock-down and control vectors(si-circFOXO3,si-NC)were constructed,and designed as the overexpression group and control group,knock-down and control group,respectively.Cell counting kit-8(CCK-8)was used to detect the proliferation of gastric cancer cells,and Transwell migration and invasion assay was used to detect the ability of migration and invasion respectively.The measurement data between groups were compared by T test,and the count data were compared by chi-square test.Results The positive expression rate of circFOXO3 in gastric cancer tissues(29/48,60.42%)was significantly higher than that in adjacent gastric normal tissues(16/48,33.33%)(χ2=7.069,P<0.05),and it was positively correlated with the stage of tissue differentiation(χ2=5.976,P<0.05),infiltration depth(χ2=4.409,P<0.05)and TNM stage(χ2=5.581,P<0.05).The results of CCK-8 and Transwell migration and invasion assays showed that overexpression of circFOXO3 significantly increased the proliferation(24 h:0.95±0.08;48 h:1.76±0.16),invasion[(157±11)cells]and migration[(186±14)cells]ability of AGS cells as compared with the control group[24 h:0.58±0.04,t=7.165,P<0.05;48 h:0.88±0.07,t=8.728,P<0.05;invasion:(85±7)cells,t=9.565,P<0.05;migration:(94±8)cells,t=9.882,P<0.05],while knockdown of circFOXO3 significantly decreased the proliferation(24 h:0.46±0.02,48 h:0.73±0.05),invasion[(53±4)cells]and migration[(65±5)cells]ability of SGC-7901 cells as compared with the control group[24 h:0.63±0.04,t=6.584,P<0.05;48 h:1.34±0.14,t=7.107,P<0.05;invasion:(96±8)cells,t=8.327,P<0.051;migration:(114±9)cells,t=8.243,P<0.05].Conclusion CircFOXO3 is highly expressed in gastric cancer tissues,and can promote the proliferation,migration and invasion of gastric cancer cells.