摘要
目的通过基因干扰长链非编码RNA(lncRNAs)SPRY4-IT1,探究其对胆囊癌(GBC)细胞GBC-SD肿瘤干细胞样特性及线粒体膜电位变化的影响。方法设计合成SPRY4-IT1 shRNA,将shRNA-SPRY4-IT1转染GBC-SD细胞,RT-PCR检测SPRY4-IT1的表达水平;BrdU染色检测细胞增殖;流式检测细胞凋亡;肿瘤细胞成球实验检测肿瘤干细胞样特性;流式检测线粒体膜电位的变化;Western blot检测肿瘤干样特性标志物以及线粒体损伤标志物的蛋白表达。结果将shRNA-SPRY4-IT1转染GBC-SD细胞,SPRY4-IT1的表达水平降低。干扰SPRY4-IT1表达抑制GBC-SD细胞增殖;提高线粒体膜电位,促进GBC-SD细胞凋亡;降低GBC干细胞样特性;抑制SOX2,OCT4,CD44,c-Myc和Bcl-2的表达以及促进cleaved Caspase-3,cleaved Caspase-9和Bax的表达。结论基因干扰lncRNAs SPRY4-IT1能降低GBC细胞GBC-SD肿瘤干细胞特性,提高线粒体膜电位,促进GBC细胞GBC-SD的凋亡。
Objective To investigated the effects of gene-interfering long-chain non-coding RNA(lncRNA)SPRY4-IT1 on stem cell-like characteristics and mitochondrial membrane potential changes in gallbladder cancer cell line GBC-SD.Methods shRNA-SPRY4-IT1 shRNA was designed and synthesized.shRNA-SPRY4-IT1 was transfected into GBC-SD cells,and the expression level of SPRY4-IT1 was detected by RT-PCR.BrdU staining was used to detect cell proliferation.Cell apoptosis was detected by flow cytometry.Tumor cell pelleting assay was used to detect the characteristics of tumor stem cells.The changes of mitochondrial membrane potential were detected by flow cytometry.Western blot was used to detect the protein expression of dry tumor characteristic markers and mitochondrial damage markers.Results shRNA-SPRY4-IT1 was transfected into GBC-SD cells,the expression level of SPRY4-IT1 decreased.Interference with SPRY4-IT1 inhibited proliferation of GBC-SD cells,improved mitochondrial membrane potential and promoted apoptosis of GBC-SD cells,reduced stem cell-like properties of gallbladder cancer,inhibited the expression of SOX2,OCT4,CD44,c-Myc,Bcl-2 and promoted the expression of cleaved Caspase-3,cleaved Caspase-9,Bax.Conclusion Gene interference with lncRNA SPRY4-IT1 can reduce the characteristics of GBC-SD cancer stem cells in gallbladder cancer cells,increase mitochondrial membrane potential,and promote apoptosis of gallbladder cancer cell line GBC-SD.
作者
李辰
王庆治
歧红阳
毛建娜
魏小娟
王鹏立
董志超
Li Chen;Wang Qingzhi;Qi Hongyang(Dept of Gastroenterology,Xinxiang Central Hospital,Xinxiang 453000;Dept of Gastroenterology,The Third Affiliated Hospital of Xinxiang Medical College,Xinxiang 453000)
出处
《安徽医科大学学报》
CAS
北大核心
2021年第3期369-374,共6页
Acta Universitatis Medicinalis Anhui
基金
河南省医学科技攻关计划项目(编号:2018020934)。
