CAJ | 학술논문

目的探讨微小RNA(microRNA,miR) 1283异常表达对滋养细胞系HTR-8/SVneo细胞浸润、增殖和凋亡的影响.方法应用miR-1283类似物(as-miR-1283)和miR 1283抑制物(anti-miR-1283)分别瞬时转染HTR-8/SVneo细胞,以转染无功能序列作为阴性对照组.荧光定量-聚合酶链反应技术检测各组细胞miR-1283的表达量,3(4,5)-二甲基噻唑-(2,5)-二苯基溴化四氮唑蓝法检测转染24、48和72 h的细胞增殖抑制率.采用流式细胞技术检测转染48 h时的细胞凋亡率.Transwell小室浸润实验观察转染24 h时各组细胞的浸润能力.组间比较采用单因素方差分析,两两检验采用Bonferroni法.结果 as-miR-1283组miR 1283的表达水平明显高于阴性对照组,差异有统计学意义(14.85±0.57与7.08±0.20,P＜0.01).转染24、48和72 h,as miR-1283组细胞增殖抑制率分别为(8.04±0.27)％、(32.47±0.24)％和(9.23±0.17)％,高于阴性对照组[分别为(0.72±0.06)％、(1.17±0.04)％和(0.53±0.05)％],差异均有统计学意义(P均＜0.01).as miR-1283组细胞凋亡率明显高于阴性对照组,差异有统计学意义[(16.33±0.40)％与(9.39±0.58)％,P＜0.01].as-miR-1283组平均穿膜细胞数少于阴性对照组,差异有统计学意义[(7.25±1.83)个与(16.33±2.08)个,P＜0.01].anti-miR-1283组miR-1283的表达水平、细胞凋亡率和穿膜细胞数与阴性对照组比较差异均无统计学意义(P均＞0.05).结论 MiR-1283高表达抑制HTR-8/SVneo细胞的浸润、增殖,促进细胞凋亡.
목적 탐토미소RNA(microRNA,miR) 1283이상표체대자양세포계HTR-8/SVneo세포침윤、증식화조망적영향.방법 응용miR-1283유사물(as-miR-1283)화miR 1283억제물(anti-miR-1283)분별순시전염HTR-8/SVneo세포,이전염무공능서렬작위음성대조조.형광정량-취합매련반응기술검측각조세포miR-1283적표체량,3(4,5)-이갑기새서-(2,5)-이분기추화사담서람법검측전염24、48화72 h적세포증식억제솔.채용류식세포기술검측전염48 h시적세포조망솔.Transwell소실침윤실험관찰전염24 h시각조세포적침윤능력.조간비교채용단인소방차분석,량량검험채용Bonferroni법.결과 as-miR-1283조miR 1283적표체수평명현고우음성대조조,차이유통계학의의(14.85±0.57여7.08±0.20,P＜0.01).전염24、48화72 h,as miR-1283조세포증식억제솔분별위(8.04±0.27)％、(32.47±0.24)％화(9.23±0.17)％,고우음성대조조[분별위(0.72±0.06)％、(1.17±0.04)％화(0.53±0.05)％],차이균유통계학의의(P균＜0.01).as miR-1283조세포조망솔명현고우음성대조조,차이유통계학의의[(16.33±0.40)％여(9.39±0.58)％,P＜0.01].as-miR-1283조평균천막세포수소우음성대조조,차이유통계학의의[(7.25±1.83)개여(16.33±2.08)개,P＜0.01].anti-miR-1283조miR-1283적표체수평、세포조망솔화천막세포수여음성대조조비교차이균무통계학의의(P균＞0.05).결론 MiR-1283고표체억제HTR-8/SVneo세포적침윤、증식,촉진세포조망.
Objective To explore the effects of microRNA(miR)-1283 on invasion,proliferation and apoptosis of HTR 8/SVneo cell line derived from human-trophoblast cells.Methods HTR-8/SVneo cells were divided into three groups,as-miR 1283 group was transfected with miR 1283 analogues,anti-miR-1283 group was transfected with miR-1283 inhibitors,and negative control group was transfected with non functional sequence.The levels of miR 1283 expression were detected by fluorescence quantitative polymerase chain reaction at 24 hours after transfection.The cell proliferation was measured by 3-(4,5)-dimethylthiazol-2-yl-(2,5)-diphenyl tetrazolium bromide assay at 24,48 and 72 hours after transfection.Apoptosis was evaluated by propidium iodide staining and flow cytometry at 48 hours after transfection.Transwell experiments were performed to evaluate cellular invasion abilities at 24 hours after transfection.Analysis of variance and Bonferroni method were applied as statistical methods.Results The level of miR 1283 in as miR 1283 group was higher than that in the negative control group with statistically significant difference (14.85±0.57 vs 7.08±0.20,P＜0.01).At 24,48 and 72 hours after transfection,the inhibitory rate of cell proliferation in as-miR-1283 group was (8.04 ± 0.27) ％,(32.47 ± 0.24) ％ and (9.23± 0.17) ％,higher than those in the negative control group [(0.72 ± 0.06) ％,(1.17 ± 0.04) ％ and (0.53 ± 0.05) ％] (P ＜ 0.01,respectively).Cell apoptosis rate was higher in as-miR 1283 group than in negative control group [(16.33 ± 0.40) ％ vs (9.39± 0.58) ％,P＜0.01].The transmembrane cell number was lower in as-miR-1283 group as comparing with the negative control group (7.25 ± 1.83 vs 16.33 ± 2.08,P＜0.01).miR-1283 expression,apoptosis and transmembrane cell number in anti-miR-1283 group had no statistical difference as compared to the negative control group (all P ＞ 0.05).Conclusions Up-regulated levels of miR-1283 could inhibit HTR-8/SVneo cell proliferation and invasion,but promote the cell apoptosis.