目的 探讨LncRNA OIP5-AS1对食管鳞状细胞癌(ESCC)细胞增殖和凋亡的影响及其机制.方法 将TE-1细胞随机分为对照组(正常培养)、si-NC组(转染 si-NC)、si-OIP5-AS1 组(转染 si-OIP5-AS1)、si-OIP5-AS1+NC inhibitor 组(转染 si-OIP5-AS1、NC inhibitor)、si-OIP5-AS1+miR-129 inhibitor组(转染 si-OIP5-AS1、miR-129 inhibitor)、NC mimic 组(转染 NC mimic)、miR-129 mimic 组(转染 miR-129 mimic)、miR-129 mimic+Vector 组(转染miR-129 mimic、Vector)、miR-129 mimic+KRAS 组[转染 miR-129 mimic、Kirsten大鼠肉瘤病毒癌基因(KRAS)].用实时荧光定量聚合酶链反应法检测各组细胞中OIP5-AS1、miR-129的表达水平,用蛋白质印迹实验检测细胞中KRAS、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、E-钙黏蛋白(E-cadherin)蛋白的表达水平,用细胞计数试剂盒-8(CCK-8)检测细胞的增殖情况,用原位末端标记(TUNEL)实验检测各组细胞的凋亡情况.结果 对照组、si-NC组、si-OIP5-AS1 组、si-OIP5-AS1+NC inhibitor 组和 si-OIP5-AS1+miR-129 inhibitor 组细胞 OIP5-AS1 表达量分别为 1.00±0.13、0.98±0.12、0.25±0.04、0.25±0.02 和 0.89±0.08,miR-129 表达量分别为1.00±0.15、0.97±0.07、2.06±0.17、2.04±0.11 和 1.22±0.15,72 h 光密度(OD)分别为 1.16±0.12、1.11±0.09、0.58±0.03、0.58±0.05 和 0.94±0.10.NC mimic、miR-129 mimic、miR-129 mimic+Vector、miR-129 mimic+KRAS 组的KRAS蛋白相对表达水平分别为1.08±0.07、0.41±0.06、0.40±0.06和1.03±0.10,72 h OD 值分别为 1.17±0.10、0.59±0.03、0.60±0.04 和0.90±0.05.si-NC 组的上述指标与 si-OIP5-AS1 组比较,si-OIP5-AS1+NC inhibitor 组 的上述指标与 si-OIP5-AS1+miR-129 inhibitor 组 比较,NC mimic组的上述指标与 miR-129 mimic 组比较,miR-129 mimic+Vector 组的上述指标与miR-129 mimic+KRAS组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 OIP5-AS1可通过调节miR-129靶向KRAS,促进ESCC细胞增殖和上皮间质转化.
Objective To investigate the effects and mechanisms of lncRNA OIP5-AS1 on cell proliferation and apoptosis in esophageal squamous cell carcinoma(ESCC).Methods TE-1 cells were randomly divided into control(normal culture),si-NC(transfected with si-NC),si-OIP5-AS1(transfected with si-OIP5-AS1),si-OIP5-AS1+NC inhibitor(transfected with si-OIP5-AS1,NC inhibitor),si-OIP5-AS1+miR-129 inhibitor(transfected with si-OIP5-ASS1,miR-129 inhibitor),NC mimic(transfected with NC mimic),miR-129 mimic(transfected with miR-129 mimic),miR-129 mimic+Vector(transfected with miR-129 mimic,Vector),miR-129 mimic+KRAS[transfected with miR-129 mimic,Kirsten rat sarcoma virus oncogene(KRAS)]groups.The expression of OIP5-AS1 and miR-129 in each group was detected by real-time fluorescence quantitative polymerase chain reaction assay.The expression levels of KRAS,N-cadherin,Vimentin and E-cadherin in cells were detected by Western blot assay.Cell proliferation was detected by cell counting kit-8(CCK-8),and apoptosis was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL)assay.Results The expression levels of OIP5-AS1 in cells of control,si-NC,si-OIP5-AS1,si-OIP5-AS1+NC inhibitor,si-OIP5-AS1+miR-129 inhibitor groups were 1.00±0.13,0.98±0.12,0.25±0.04,0.25±0.02 and 0.89±0.08;the expression levels of miR-129 were 1.00±0.15,0.97±0.07,2.06±0.17,2.04±0.11 and 1.22±0.15;72 h absorbance values(OD)were 1.16±0.12,1.11±0.09,0.58±0.03,0.58±0.05 and 0.94±0.10.The KRAS protein expression levels of NC mimic,miR-129 mimic,miR-129 mimic+Vector,and miR-129 mimic+KRAS groups were 1.08±0.07,0.41±0.06,0.40±0.06,1.03±0.10;the 72 h OD values were 1.17±0.10,0.59±0.03,0.60±0.04 and 0.90±0.05,respectively.si-NC group was compared with si-OIP5-AS1 group,si-OIP5-AS1+NC inhibitor group was compared with si-OIP5-AS1+miR-129 inhibitor group,NC mimic group was compared with miR-129 mimic group,miR-129 mimic+Vector group was compared with miR-129 mimic+KRAS group,the differences of the above indexes were statistically significant(all P＜0.05).Conclusion OIP5-AS1 can promote ESCC cell proliferation and epithelial mesenchymal transformation by regulating miR-129 targeting KRAS.
许俊凯;刘剑雄;陈奇松;赵云辉
莆田学院附属医院放疗科,福建莆田 351100莆田学院附属医院消化内科,福建莆田 351100
药学
长链非编码RNA OIP5-AS1;食管癌;Kirsten大鼠肉瘤病毒癌基因;微小RNA-129;增殖;上皮间质转化
long non-coding RNA OIP5-AS1;esophageal cancer;Kirsten rat sarcoma virus oncogene;microrna-129;proliferation;epithelial mesenchymal transformation
《中国临床药理学杂志》 2024 (011)
1573-1577 / 5
福建省教育厅中青年教师教育科研基金资助项目(JAT200531)
10.13699/j.cnki.1001-6821.2024.11.007
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