Lei Chang-Jiang, Yao Chun, Li De-Ke, Long Zhi-Xiong, Li Yuan, Tao Dan, Liou Yan-Ping, Zhang Jiang-Zhou, Liu Ning
Tumor Laboratory, The Fifth Hospital of Wuhan, Wuhan, 430050, Hubei Province, China.
Wuhan Hematology Institute, Wuhan, 430050, Hubei Province, China.
Asian Pac J Trop Med. 2016 Sep;9(9):898-902. doi: 10.1016/j.apjtm.2016.07.015. Epub 2016 Jul 26.
To investigate the effects of co-transfection of miR-520c-3p and miR-132 on proliferation and apoptosis of hepatocellular carcinoma Huh7.
Hepatocellular carcinoma Huh7 was cultured in vitro and lipidosome was used to transfect miR-520c-3p and miR-132, respectively or together. The effects of transfection of miR-520c-3p and miR-132 on proliferation and apoptosis of Huh7 were detected by CCK8 and Annexin V staining and flow cytometry, and the expression level of the targeted gene of over-expressed miR-520c-3p and miR-132 was determined by Western blot and realtime PCR.
Compared with the control group, the proliferation ability of Huh7 of the single transfected and co-transfected miR-520c-3p and miR-132 decreased significantly, and the apoptosis ratio increased distinctly (P < 0.05). Besides, the effect of the co-transfection group was better than that of the single transfection group. The protein levels of GPC3 (Glypican-3) and YAP (Yes-associated protein), the target genes transfected only by miR-520c-3p and miR-132, respectively, reduced obviously (P < 0.05), which was similar with the co-infected cells, but cells transfected by miR-132 only showed a decrease of YAP.
The co-transfection of miR-520c-3p and miR-132 can target-regulate the expression of GPC3 and YAP, enhance the exhibition effect on proliferation of hepatocellular carcinoma Huh7 and induce cell apoptosis synergistically.
探讨共转染miR-520c-3p和miR-132对肝癌Huh7细胞增殖和凋亡的影响。
体外培养肝癌Huh7细胞，分别或共同使用脂质体转染miR-520c-3p和miR-132。采用CCK8法、Annexin V染色及流式细胞术检测miR-520c-3p和miR-132转染对Huh7细胞增殖和凋亡的影响，采用蛋白质印迹法和实时荧光定量PCR法检测过表达miR-520c-3p和miR-132的靶基因表达水平。
与对照组相比，单独转染及共转染miR-520c-3p和miR-132的Huh7细胞增殖能力显著降低，凋亡率明显升高（P<0.05）。此外，共转染组的效果优于单独转染组。仅被miR-520c-3p和miR-132转染的靶基因GPC3（磷脂酰肌醇蛋白聚糖-3）和YAP（Yes相关蛋白）的蛋白水平明显降低（P<0.05），这与共感染细胞相似，但仅转染miR-132的细胞YAP水平降低。
共转染miR-520c-3p和miR-132可靶向调控GPC3和YAP的表达，增强对肝癌Huh7细胞增殖的抑制作用并协同诱导细胞凋亡。
