多囊卵巢综合征患者血清外泌体中miR-184的表达水平及其对卵巢颗粒细胞增殖的影响

目的研究多囊卵巢综合征(PCOS)患者血清外泌体中miR-184的表达水平及其对卵巢细胞颗粒增殖的影响。方法回顾性选取2018年2月至2020年4月期间盘锦辽油宝石花医院收治的60例PCOS患者作为PCOS组,另纳入同期30名健康成年女性作为对照组,提取2组受试者血清外泌体。应用实时荧光定量聚合酶链式反应(qRT-PCR)法检测2组受试者血清外泌体中及人卵巢颗粒细胞KGN和人正常卵巢上皮细胞IOSE80中的miR-184的相对表达。转染miR-184抑制剂的KGN细胞设为miR-184抑制剂组,转染抑制剂对照物的KGN细胞设为对照组,不做任何处理的KGN细胞设为空白组。应用MTT法检测各组KGN细胞的增殖情况,应用平板克隆实验检测各组KGN细胞的克隆形成率。结果PCOS患者血清外泌体中和KGN细胞中的miR-184的相对表达量为4.23±0.49、5.81±0.73,显著高于对照组(1.32±0.21)和IOSE80组细胞(1.19±0.15),差异有统计学意义(P<0.05)。miR-184抑制剂组KGN细胞在24、48、72、96 h的吸光度值分别为0.20±0.05、0.22±0.07、0.26±0.08、0.29±0.07,均显著低于阴性对照组(0.31±0.09、0.52±0.11、0.78±0.12、0.96±0.18)和空白对照组(0.31±0.08、0.53±0.10、0.77±0.14、0.94±0.18),克隆形成率(26.78±5.98)%显著低于阴性对照组[(45.98±4.12)%]和空白对照组[(43.56±4.34)%],差异有统计学意义(P<0.05)。结论PCOS患者血清外泌体和人卵巢颗粒细胞KGN中miR-184呈现高表达,PCOS的发病可能与miR-184促进卵巢颗粒细胞增殖有关。

Expression level of miR-184 in serum exosomes of polycystic ovary syndrome patients and its effect on the proliferation of ovarian granulosa cells

Objective To investigate the expression level of miR-184 in serum exosomes of polycystic ovary syndrome(PCOS)patients and its effect on the proliferation of ovarian granulosa cells.Methods Sixty PCOS patients addmited to Panjin liaohe oilfield GEM Flower Hospital from Febraury 2018 to April 2020 were selected as PCOS group,another 30 healthy adult female were brought into control group,the serum exosomes of two groups were extracted.QRT-PCR method was used to detected the relative expression of miR-184 in exosomes,KGN and IOSE80 cells.The KGN cells transfected with miR-184 inhibitor were set as the miR-184 inhibitor group,the KGN cells transfected with the inhibitor control substance were set as the control group,and the KGN cells without any treatment were set as the blank group.The proliferation of KGN cells was detected by MTT assay.The cell clone formation rate was detected by plate cloning assay.Results The relative expression levels of miR-184 in the serum exosomes of PCOS patients and KGN cells were 4.23±0.49 and 5.81±0.73,which were significantly higher than those of the control group(1.32±0.21)and IOSE80 cells(1.19±0.15),and the difference was statistically significant(P<0.05).The absorbance values of KGN cells in the miR-184 inhibitor group at 24,48,72 and 96 h were 0.20±0.05,0.22±0.07,0.26±0.08,0.29±0.07,respectively,which were significantly lower than those in the negative control group(0.31±0.09,0.52±0.11,0.78±0.12,0.96±0.18)and the blank control group(0.31±0.08,0.53±0.10,0.77±0.14,0.94±0.18),the clone formation rate(26.78±5.98)%was significantly lower than the negative control group[(45.98±4.12)%]and the blank control group[(43.56±4.34)%],the difference was statistically significant(P<0.05).Conclusion The expression of miR-184 in exosomes of PCOS patients and KGN cells were up-regulated,the mechanism of PCOS might be related to the promotion of proliferation of ovarian granulosa cells by miR-184.