LINC00461调节miR-1301-3p/MYH9轴对胃癌细胞增殖、凋亡及顺铂耐药性的影响  

Impacts of LINC00461 on proliferation,apoptosis,and cisplatin resistance of gastric cancer cells by regulating the miR⁃1301⁃3p/MYH9 axis

作　　者：李亚岭[1] 殷景远 LI Yaling;YIN Jingyuan(Department of Gastroenterology,Yellow River Central Hospital,Yellow River Conservancy Commission,Zhengzhou,Henan 450003,China)

机构地区：[1]黄河水利委员会黄河中心医院消化内科,河南郑州450003

出　　处：《热带医学杂志》2024年第11期1495-1500,1511,共7页Journal of Tropical Medicine

基　　金：河南省医学科技攻关计划项目(2018020875)。

摘　　要：目的探究长链非编码RNA LINC00461(LINC00461)调节微小RNA-1301-3p/肌凝蛋白重链9(miR-1301-3p/MYH9)轴对胃癌细胞增殖、凋亡及顺铂(CDDP)耐药性的影响,为胃癌的治疗寻求新靶点。方法实时荧光定量逆转录聚合酶链式反应(RT-qPCR)检测正常胃黏膜细胞GES-1、胃癌细胞HGC-27、胃癌顺铂耐药细胞株HGC-27/CDDP中LINC00461、miR-1301-3p及MYH9表达情况;将胃癌顺铂耐药细胞HGC-27/CDDP分为对照组(Control组)、si-NC组、si-LINC00461组、si-LINC00461+anti-NC组、si-LINC00461+anti-miR-1301-3p组;RT-qPCR法检测各组HGC-27/CDDP细胞中LINC00461、miR-1301-3p及MYH9表达情况;Edu实验检测细胞增殖;Annexin V-FITHCA/PI检测细胞凋亡;Western blot法检测细胞增殖、凋亡相关蛋白表达;双荧光素酶实验检测LINC00461、MYH9分别与miR-1301-3p的靶向关系。结果在HGC-27/CDDP细胞中LINC00461、MYH9 mRNA高表达,miR-1301-3p低表达,与HGC-27、GES-1细胞比较,差异均有统计学意义(F=29.503、31.153、87.750,P均<0.05)。网站预测及双荧光素酶实验显示,LINC00461、MYH9与miR-1301-3p之间具有靶向关系;LINC00461-WT+miR-1301-3p mimic组较LINC00461-WT+miR-NC组相对荧光素酶活性降低,差异有统计学意义(t=9.202,P<0.05);MYH9-WT+miR-1301-3p mimic组较MYH9-WT+miR-NC组相对荧光素酶活性降低,差异有统计学意义(t=10.944,P<0.05)。与si-NC组相比,siLINC00461组LINC00461、MYH9 mRNA水平、Edu阳性率及CyclinD1、Ki67表达降低(t=7.351、9.992、15.424、9.640、11.192),miR-1301-3p水平、细胞凋亡率及Bax、Caspase-3表达升高(t=8.104、34.304、6.341、7.473),差异均有统计学意义(P均<0.05)。与si-LINC00461+anti-NC组相比,si-LINC00461+anti-miR-1301-3p组MYH9 mRNA水平、Edu阳性率及CyclinD1、Ki67表达升高(t=8.568、13.630、7.851、8.828),miR-1301-3p水平、细胞凋亡率及Bax、Caspase-3表达降低(t=6.549、22.847、4.726、4.941),差异均有统计学意义(P均<0.05)。结论沉默LINC00461可抑制胃癌细胞增殖,促进细胞Objective To investigate the impacts of long non coding RNA LINC00461(LINC00461)on the proliferation,apoptosis,and cisplatin(CDDP)resistance of gastric cancer cells by regulating the miR⁃1301⁃3p/myosin heavy chain 9(miR⁃1301⁃3p/MYH9)axis,seeking new targets for the treatment of gastric cancer.Methods The reverse transcription⁃quantitative polymerase chain reaction(RT⁃qPCR)method was applied to detect the expression of LINC00461,miR⁃1301⁃3p,and MYH9 in normal gastric mucosal cells GES⁃1,gastric cancer cells HGC⁃27,and gastric cancer cisplatin resistant cell lines HGC⁃27/CDDP.Gastric cancer cisplatin resistant cells HGC⁃27/CDDP were separated into Control group,si⁃NC group,si⁃LINC00461 group,si⁃LINC00461+anti⁃NC group,and si⁃LINC00461+anti⁃miR⁃1301⁃3p group.The RT⁃qPCR method was applied to detect the expression of LINC00461,miR⁃1301⁃3p,and MYH9 in HGC⁃27/CDDP cells in each group.Edu experiment was applied to detect cell proliferation.Annexin V⁃FITHCA/PI was applied to detect cell apoptosis.Western blot was applied to detect the expression of cell proliferation and apoptosis related proteins.Dual luciferase assay was applied to detect the targeting relationship between LINC00461 and MYH9 with miR⁃1301⁃3p,respectively.Results In HGC⁃27/CDDP cells,the expression of LINC00461 and MYH9 mRNA was high,and the expression of miR⁃1301⁃3p was low;the differences were significant when compared with HGC⁃27 and GES⁃1 cells(F=29.503,31.153,87.750;all P<0.05).Website prediction and dual luciferase experiments showed that LINC00461,MYH9,and miR⁃1301⁃3p had a targeting relationship.The relative luciferase activity of LINC00461⁃WT+miR⁃1301⁃3p mimic group was lower than that of LINC00461⁃WT+miR⁃NC group(t=9.202,P<0.05).The relative luciferase activity of MYH9⁃WT+miR⁃1301⁃3p mimic group was lower than that of MYH9⁃WT+miR⁃NC group(t=10.944,P<0.05).Compared with the si⁃NC group,the levels of LINC00461 and MYH9 mRNA,the positive rate of Edu and the

关 键 词：长链非编码RNA LINC00461 微小RNA-1301-3p 肌凝蛋白重链9 胃癌 增殖 

分 类 号：R735.2[医药卫生—肿瘤]