Chen Xiaoyu, Song Meiyi, Chen Wei, Dimitrova-Shumkovska Jasmina, Zhao Yingying, Cao Yan, Song Yang, Yang Wenzhuo, Wang Fei, Xiang Yang, Yang Changqing
Division of Gastroenterology and Hepatology, Digestive Disease Institute, Tongji Hospital, Tongji University School of Medicine, Shanghai, China (mainland).
Emergency Department, Tongji Hospital, Tongji University School of Medicine, Shanghai, China (mainland).
Med Sci Monit. 2016 Jan 8;22:83-91. doi: 10.12659/MSM.896157.
Multiple microRNAs (miRNAs, miRs), including miR-21, have been documented to be critical regulators of liver regeneration, but the mechanism underlying their roles in hepatocyte proliferation and cell cycle progression is still far from understood.
MATERIAL/METHODS: miR-21 levels were determined using qRT-PCRs in mouse livers at 48 h after 70% partial hepatectomy (PH-48 h). Cell proliferation was determined by use of a cell-counting kit-8 (CCK-8), EdU incorporation staining, and flow cytometry. Phosphatase and tensin homolog (PTEN) expressions were determined using qRT-PCR and Western blot analysis. PTEN siRNA was used to perform the rescue experiment.
A marked upregulation of miR-21 was observed in mouse livers at 48 h after 70% partial hepatectomy (PH-48 h) compared to 0 h after PH (PH-0 h). Overexpression of miR-21 was associated with increased proliferation and a rapid G1-to-S phase transition of the cell cycle in BNL CL.2 normal liver cells in vitro. In addition, we showed that PTEN expression was inversely correlated with miR-21 in BNL CL.2 cells and demonstrated that PTEN expression is lower in mouse livers at PH-48 h. Moreover, the presence of PTEN siRNA significantly abolished the suppressive effect of miR-21 inhibitor on hepatocyte proliferation.
miR-21 overexpression contributes to liver regeneration and hepatocyte proliferation by targeting PTEN. Upregulation of miR-21 might be a useful therapeutic strategy to promote liver regeneration.
多种微小RNA（miRNA，miR），包括miR-21，已被证明是肝脏再生的关键调节因子，但其在肝细胞增殖和细胞周期进程中发挥作用的机制仍远未明确。
材料/方法：采用qRT-PCR法测定70%部分肝切除术后48小时（PH-48 h）小鼠肝脏中的miR-21水平。使用细胞计数试剂盒-8（CCK-8）、EdU掺入染色和流式细胞术测定细胞增殖。采用qRT-PCR和蛋白质免疫印迹分析测定磷酸酶和张力蛋白同源物（PTEN）的表达。使用PTEN siRNA进行挽救实验。
与部分肝切除术后0小时（PH-0 h）相比，在70%部分肝切除术后48小时（PH-48 h）的小鼠肝脏中观察到miR-21显著上调。在体外，miR-21的过表达与BNL CL.2正常肝细胞的增殖增加和细胞周期从G1期到S期的快速转变相关。此外，我们发现PTEN的表达与BNL CL.2细胞中的miR-21呈负相关，并证明在PH-48 h的小鼠肝脏中PTEN表达较低。此外，PTEN siRNA的存在显著消除了miR-21抑制剂对肝细胞增殖的抑制作用。
miR-21的过表达通过靶向PTEN促进肝脏再生和肝细胞增殖。miR-21的上调可能是促进肝脏再生的一种有用的治疗策略。
