干扰hsa_circRNA_002178基因抑制卵巢癌细胞的恶性生物学行为  

hsa_circRNA_002178 Gene Interference Inhibits Malignant Biological Behavior of Ovarian Cancer Cells

作　　者：彭胡 唐克彬 罗婷 尹帮朝 周剑 PENG Hu;TANG Kebin;LUO Ting;YIN Bangchao;ZHOU Jian(Department of Medical Laboratory,Luzhou Traditional Chinese Medicine Hospital,Luzhou,Sichuan,646000,China;Department of Pathology,Yibin Sixth People’s Hospital,Yibin,Sichuan,644000,China;Department of Pathology,Southwest Medical University,Luzhou,Sichuan,646000,China)

机构地区：[1]泸州市中医医院医学检验科,四川省泸州市646000 [2]宜宾市第六人民医院病理科,四川省宜宾市644000 [3]西南医科大学病理科,四川首泸州市646000

出　　处：《医学分子生物学杂志》2024年第4期347-353,360,共8页Journal of Medical Molecular Biology

基　　金：泸州市科技计划项目(No.2021-S-46)。

摘　　要：目的研究干扰circRNA_002178基因对卵巢癌细胞恶性生物学行为的影响。方法将人卵巢癌SKOV3细胞分为对照组、短发夹RNA-阴性对照(shRNA-NC)组、circRNA_002178-shRNA1和circRNA_002178-shRNA2组。采用实时荧光定量聚合酶链反应(RT-PCR)检测各组细胞中circRNA_002178的表达;流式检测细胞凋亡、干细胞阳性(CD44,CD133)细胞株及线粒体膜电位的变化;干细胞成球实验检测干细胞成球直径和成球率;Transwell检测细胞侵袭,蛋白印迹检测相关蛋白表达。构建移植瘤模型,观察circRNA_002178基因干扰对成瘤的影响。结果与对照组比较,circRNA_002178-shRNA干扰显著促进SKOV3细胞凋亡,减少CD44、CD133阳性细胞数量、干细胞成球率、成球直径和侵袭细胞数,加剧线粒体膜电位变化,并抑制移植瘤大鼠肿瘤生长及Akt/mTOR通路的活化。结论circRNA_002178基因干扰可抑制卵巢癌细胞的恶性生物学行为,其机制可能与抑制Akt/mTOR通路有关。Objective To study the effect of circRNA_002178 gene interference on malignant biological behavior of ovarian cancer cells.Methods Human ovarian cancer SKOV3 cells were divided into 4 groups:control group,short hairpin RNA negative control(shRNA-NC)group,circRNA_002178-shRNA 1 group and circRNA_002178-shRNA 2 group.The expression level of circRNA_002178 was detected by real-time fluorescence quantitative polymerase chain reaction(RTPCR).Flow cytometry was used to detect the cell apoptosis and mitochondrial membrane potential and to select the stem cell positive(CD44,CD133)cell lines.The stem cell spheroidization test was used to detect the sphere formation rate and the diameter of spheres.Transwell assay was used to detect the cell invasion.Western blotting was used to detect the expression levels of the proteins.The transplanted tumor model was constructed to observe the effect of circRNA_002178 gene interference on tumor formation.Results CircRNA_002178 interference significantly promoted the apoptosis of SKOV3 cells,decreased the number of CD44 and CD133 positive cells,the rate of sphere formation,the diameter of spheres and the number of invasive cells,intensified the changes of mitochondrial membrane potential,and inhibited tumor growth and Akt/mTOR pathway activation in transplanted tumor rats.Conclusion CircRNA_002178 interference can inhibit the malignant biological behavior of ovarian cancer cells.The mechanism may be related to the inhibitory effect on Akt/mTOR pathway.

关 键 词：circRNA_002178 CD133 线粒体膜电位 细胞凋亡 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]