摘要
目的观察重编程调节因子(ROR)在胃癌组织中的表达,探讨ROR对胃癌细胞生物学行为的影响。方法实时定量反转录聚合酶链反应(RT-qPCR)检测50例胃癌及其配对正常组织中ROR的表达水平;利用基因过表达技术构建ROR过表达载体,慢病毒法转染MGC-803、SGC-7901细胞,RT-qPCR检测过表达效果;实验分空白对照组(NC)、空载对照组(LV5-NC)及过表达组(LV5-ROR):分别利用细胞计数实验(CCK-8)、流式细胞术和Transwell迁移及侵袭实验检测过表达ROR对胃癌细胞增殖、迁移及侵袭的影响。结果50例胃癌组织中ROR的表达量明显低于正常对照组织(P<0.01);CCK-8实验结果显示胃癌细胞株过表达ROR后,与空白对照组和空载对照组比较,在450 nm处的吸光度(A)值差异无统计学意义(P>0.05);Transwell迁移实验结果显示过表达组胃癌细胞的穿膜数(336.33±34.93、201.67±45.35)较空载对照组(841.67±38.19、503.34±47.00)及空白对照组(826.00±35.20、520.25±39.50)明显减少(P<0.01),侵袭实验结果也显示过表达后细胞穿膜数(253.33±41.63、126.67±25.17)较空载对照组(757.67±52.73、311.34±36.23)及空白对照组(762±48.53、340±28.225)明显减少(P<0.01)。结论ROR在胃癌组织中表达降低,过表达ROR后胃癌细胞迁移及侵袭能力减弱,但对细胞增殖无明显影响,表明ROR通过抑制胃癌细胞的迁移及侵袭能力阻止胃癌的发生与进展。
Objective To explore the expression of regulator of reprogramming (ROR) in human gastric cancer tissues and investigate the effects of ROR on biological behaviors in vitro. Methods Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect the expression of ROR in 50 gastric cancer tissues and its paired normal tissues. To further explore its biological function, over-expression approach was used to up ROR expression. Normal control group (NC), plasmid control group (LV5-NC) and over-expression group (LV5-ROR)were transfected to in MGC-803, SGC-7901, and the transfection efficiency was evaluated by RT-qPCR. Cell counting kit-8 (CCK-8), transwell migration and invasion assays were performed to detect the biological effects of ROR in gastric cancer cells after transfection. Results The level of ROR was significantly down-regulated in gastric cancer compared with paired normal tissues samples (P<0.01). CCK-8 assay indicated that there is no difference between LV5-ROR, LV5-NC and NC (P>0.05). Transwell migration assay showed that the number of cells passing the membrance in LV5-ROR (336.33±34.93, 201.67±45.35) were decreased significantly than LV5-NC(841.67±38.19, 605.25±45.00)and NC 790.58±35.68, 620.59±49.00)(P<0.01). As is also in invasion assay, we further discovered the number of cells passing the membrance in LV5-ROR (253.33±41.63, 290.56±35.36) were lower than LV5-NC (757.67±52.73, 680.56±32.93) and NC (790.35±51.73, 780.35±32.85)(P<0.01). Conclusion Our results suggested that ROR was significantly down-regulated in gastric cancer tissues. These studies indicated over-expression of ROR in gastric cancer cells could suppressed invasion and migration, while had little effect on cell proliferation. ROR could prevent carcinogenesis and progression by inhibiting migration and invasion.
作者
朱长明
王涛
高华
徐垚
闫峰
Zhu Changming;Wang Tao;Gao Hua;Xu Yao;Yan Feng(Department of Gastrointestinal Surgery, the Affiliated Zhongshan Hospital, Xiamen University, Xiamen 361004 , China;Department of General Surgery, the Third People Hospital of Hubei Province, Wuhan 430015, China;Department of Gastrointestinal Surgery, the Affiliated Zhongs han Hospital, Xiamen University ,Xiamen 361004, China , Institute of Gastrointestinal Oncology , Medical College of Xiamen University, Xiamen, Fujian, 361004, China;Department of Gastrointestinal Tract, the People's Hospital of Zhangjiajie, Zhangjiajie 427000, China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第3期411-413,共3页
Chinese Journal of Experimental Surgery
基金
福建省自然科学基金(2015J01522).
关键词
胃癌
长链非编码RNA
重编程调节因子
Gastric cancer
Long non-coding RNA
Regulator of reprogramming
