miRNA-519d对卵巢癌的生长抑制作用及其对XIAP表达的影响

Effects of MicroRNA-519d on the Growth and Expression of XIAP of Ovarian Cancer Cell Line

目的探讨MicroRNA-519d(miRNA-519d)对卵巢癌细胞生长抑制的作用及其对XIAP表达的影响。方法对人卵巢癌细胞系OVCAR3瞬时转染miR-519d mimic(实验组)和NC(阴性对照组),不进行任何转染为空白对照组,转染48 h后进行MTT实验、流式细胞实验、Transwell侵袭实验,观察卵巢癌细胞生长抑制状况,WB实验测定XIAP的表达变化。结果定量Real-time PCR检测显示:miR-519d的表达水平均较对阴性对照组与空白对照组有明显的增高(P<0.05)。MTT检测显示:上调OVCAR3细胞内miR-519d水平后,细胞增殖抑制率为(29.81±8.24)%,而阴性照组与空白对照组分别为(2.49±0.28)%和(1.98±0.34)%,对比差异有统计学意义(P<0.05)。流式细胞术检测显示实验组OVCAR3细胞凋亡率为(8.33±1.39)%,而阴性对照组与空白对照组分别为(2.13±2.11)%和(1.89±1.08)%,对比差异有统计学意义(P<0.05)。Transwell侵袭实验显示:转染miR-519dmimics后实验组、阴性对照组、空白对照组的三组的细胞转移能力对比无明显差异(P>0.05)。Western实验结果显示:处理后实验组的XIAP蛋白相对表达量为(0.98±0.34),阴性对照组为(7.54±1.22),空白对照组为(7.67±1.32),对比差异有统计学意义(P<0.05)。结论增强miR-519d的表达能对卵巢癌细胞的增殖起抑制作用,并促进其凋亡,而对卵巢癌细胞的转移没有影响,能抑制XIAP的表达,表明miR-519d在卵巢癌的生长中起到一定的抑制作用。

Objective To study the effects of MicroRNA-519d （miR-519d） on the the growth and expression of XIAP of ovarian cancer cell line. Methods The human ovarian cancer cell lineOVCAR3 were transfected to miR-519d mimic （ experi- mental group） and NC （ negative control group） , no transfection were the blank control group, after transfection of 48 h were given MTF assay,flow cytometry assay and Transwe｜l invasion experiments, observed the growth of ovarian cancer cell. The expression of XIAP was determined by WB assay. Results quantitative PCR Real-time test showed that the expression level of miR-519 dwere significantly higher than that of negative group and blank control group （ P 〈 0.05 ）. MTT showed that the cell proliferation inhibition rate in the experimental group was （29.81 ± 8.24） %, and the negative control group and blank control group were （2.49 ± 0.28） % and （ 1.98 ± 0.34） %, compared were statistically significant difference （ P 〈 0.05 ）. Flow cytometry showed that the apoptosis rate of experimental group was （8.33 ± 1.39）% ,while the negative group and the blank control group were （2.13 ± 2.11 ） % and （ 1.89 ± 1.08 ） % respectively. The Transwell invasion experiment showed that there were no significant difference compared among the three groups（ P 〉 0. 05 ）. The results of Western showed that the relative expression of XIAP protein in the experimental group was （0.98 ± 0.34）, and the negative control group was （7.52 ± 1.22）, with a blank control group of （7.67 ± 1.32）, and the difference was statistically significant （P 〈 0.05 ）. Conclusion Transient transfection of mimics miR-519d in the ovarian cancer cell line can inhibit the cell growth and promote its apoptosis but has no effect on the metastasis,it can inhibit the expression of XIAP,which indicates that miR-519d might inhibit the proliferation of ovarian cancer cells at the post transcrip- tional level.