妊娠糖尿病患者血清microRNA-21调节Bcl-2/Caspase-9对滋养层细胞生物学行为的影响

目的 探讨妊娠糖尿病（GDM）患者血清microRNA-21（miR-21）表达变化及其对滋养层细胞生物学行为的作用机制。方法 选取2019年1月—2020年10月三亚市妇幼保健院收治的66例GDM孕妇及同期体检且孕周相近的38例健康孕妇作为研究对象。采用实时荧光定量聚合酶链反应（qRT-PCR）检测血清miR-21 mRNA表达水平，microRNA转染JAR细胞系构建Control、miR-21 NC、miR-21、miR-21 inhibitor NC和miR-21 inhibitor组细胞。采用CCK-8法检测细胞活力，Brdu检测细胞增殖，Matrigel侵袭实验检测细胞侵袭，TUNEL法检测细胞凋亡；采用Western blotting检测Bcl-2、Bax、Cleaved-Caspase 9、E-cadherin及Vimentin蛋白表达。结果 研究组患者miR-21 mRNA相对表达量及FPG、HbA1c水平较对照组高（P <0.05）。miR-21组细胞miR-21 mRNA相对表达量高于Control组（P <0.05），miR-21 inhibitor组细胞 mRNA相对表达量低于Control组（P <0.05）。CCK-8法检测结果显示，miR-21组细胞活力较Control组下调（P <0.05），miR-21 inhibitor组细胞活力较Control组上升（P <0.05）。miR-21组细胞增殖能力较Control组减弱（P <0.05），miR-21 inhibitor组细胞增殖能力较Control组增强（P <0.05）。miR-21组细胞凋亡率较Control组升高（P <0.05），miR-21 inhibitor组细胞凋亡率较Control组降低（P <0.05）。miR-21组的穿膜细胞数低于Control组（P <0.05），miR-21 inhibitor组的穿膜细胞数高于Control组（P <0.05）。相较于Control组，miR-21组E-cadherin表达上调，Vimentin表达下调（P <0.05），miR-21 inhibitor组E-cadherin表达下调，Vimentin表达上调（P <0.05）。相较于Control组，miR-21组Cleaved Caspased-9、Bax蛋白相对表达量上升，Bcl-2蛋白相对表达量下降（P <0.05）；miR-21 inhibitor组Caspased-9、Bax蛋白相对表达量下降，Bcl-2蛋白相对表达量上升（P <0.05）。结论 GDM孕妇存在miR-21高表达，miR-21可通过调节Bcl-2/Caspase-9介导对滋养层细胞增殖、侵袭的抑制作用，继而促进细胞凋亡。

Effect of serum microRNA-21 in pregnant women with diabetes mellitus on biological behavior of trophoblast cells by regulating Bcl-2/Caspase-9

Objective To investigate the changes of serum miRNA-21 expression in patients with gestational diabetes and its mechanism on the biological behavior of trophoblast cells.Methods From January 2019 to October 2020, 66 pregnant women with diabetes in pregnancy and 38 healthy pregnant women with similar gestational weeks who underwent physical examination at the same time were selected as the study subjects. The expression level of miR-21 in serum of the two groups was tested by qRT-PCR. The JAR cell lines were transfected with miRNA to construct control, miR-21NC, miR-21, miR-21 inhibitor NC, and miR-21 inhibitor group cells. Cell viability was tested by cell proliferation detection kit 8 (CCK-8), cell proliferation was tested by Brdu, cell invasion was tested by Matrigel invasion test, and cell apoptosis was tested by TUNEL. The expression levels of Bcl-2, Bax, Cleared-capase 9, Vimentin, and E-cadherin were tested by Western blotting.Results The expression levels of FPG, miRNA-21, and HbA1c in the study group were higher than those in the control group (P < 0.05). The expression level of miR-21 in miR-21 group and miR-21 inhibitor group was higher and lower than that in control group (P < 0.05). CCK-8 test results showed that the cell viability of miR-21 group was lower than that of control group, and that of miR-21 inhibitor group was higher than that of control group and miR-21 group (P < 0.05). Compared with control group, the proliferation ability of cells in miR-21 group and miR-21 inhibitor group decreased and increased significantly (P < 0.05). The apoptosis rate of miR-21 group and miR-21 inhibitor group was significantly higher and lower than that of control group (P < 0.05). The number of penetrating cells in miR-21 group and miR-21 inhibitor group was significantly lower than that in control group (P < 0.05). Compared with the control group, the expression of E-cadherin and Vimentin in the miR-21 group was down and up, respectively, but the expression of E-cadherin and Vimentin in the miR-21 inhibitor group was down and up, respectively, compared with the control group and the miR-21 group (P < 0.05). The expression of anti-apoptotic protein Bcl-2 decreased in miRNA-21 group and increased in miR-21 inhibitor group (P < 0.05). Compared with the control group, the expression of Bax and Cleared caspased-9 protein in the miRNA-21 group increased, while the expression of the above proteins in the miR-21 inhibitor group decreased (P < 0.05).Conclusion There is high expression of miRNA-21 in pregnant women with gestational diabetes. miRNA-21 can promote cell apoptosis by regulating the inhibitory effect of Bcl-2/Caspase-9 on trophoblast cell proliferation and invasion.