Meng Xuan, Shi Yong, Xiang Xin, Li Chonghui, Ge Xinlan, Pan Ke, Liang Yurong
Department of Hepatobiliary Surgery, Chinese PLA General Hospital, Beijing 100853, P.R. China.
Oncol Lett. 2020 Feb;19(2):1310-1316. doi: 10.3892/ol.2019.11210. Epub 2019 Dec 12.
The expression of miR-101 in carcinoma and para-carcinoma tissues of patients with liver cancer was studied. The carcinoma and para-carcinoma tissues of 67 patients with liver cancer treated in Chinese PLA General Hospital were collected, and the expression of miR-101 in carcinoma and para-carcinoma tissues was detected via reverse transcription-polymerase chain reaction (RT-PCR). The liver cancer HepG2 cell line was transfected with miR-101 mimics. Moreover, the influence of miR-101 overexpression on the proliferation of liver cancer cells was detected via Cell Counting Kit-8 assay and colony formation assay. The proportion of Ki67-positive cells in the control group (NC group) and miR-101 overexpression group (miR-101 mimics group) was detected via Ki67 staining. The proportions of cells were detected via flow cytometry, and the predicted target gene Zeste2 enhancer (EZH2) was further verified via luciferase reporter gene assay and western blotting. The miR-101 overexpression significantly inhibited the colony formation and proliferation ability of liver cancer cells (P<0.05). The proportion of Ki67-positive cells in liver cancer cells was lower in miR-101 mimics group (P<0.05). The proportion of cells in G0/G1 phase was increased in miR-101 mimics group compared with that in NC group (P<0.05). The extracellular signal-regulated kinase (ERK)1/2 phosphorylation level in liver cancer cells was obviously suppressed in miR-101 mimics group (P<0.05). Therefore, the expression level of miR-101 declines in liver cancer tissues, and the miR-101 overexpression can inhibit the proliferation of liver cancer cells. The inhibitory effect of miR-101 on the proliferation of liver cancer cells may be related to its inhibition on the mitogen-activated protein kinase (MAPK)/ERK signaling pathway, and the inhibition on the MAPK/ERK may be mediated by the targeted inhibition of miR-101 on EZH2.
研究了肝癌患者癌组织和癌旁组织中miR-101的表达情况。收集了解放军总医院收治的67例肝癌患者的癌组织和癌旁组织，通过逆转录聚合酶链反应（RT-PCR）检测癌组织和癌旁组织中miR-101的表达。用miR-101模拟物转染肝癌HepG2细胞系。此外，通过细胞计数试剂盒-8法和集落形成试验检测miR-101过表达对肝癌细胞增殖的影响。通过Ki67染色检测对照组（NC组）和miR-101过表达组（miR-101模拟物组）中Ki67阳性细胞的比例。通过流式细胞术检测细胞比例，并通过荧光素酶报告基因试验和蛋白质印迹法进一步验证预测的靶基因Zeste2增强子（EZH2）。miR-101过表达显著抑制肝癌细胞的集落形成和增殖能力（P<0.05）。miR-101模拟物组肝癌细胞中Ki67阳性细胞的比例较低（P<0.05）。与NC组相比，miR-101模拟物组中处于G0/G1期的细胞比例增加（P<0.05）。miR-101模拟物组肝癌细胞中细胞外信号调节激酶（ERK）1/2的磷酸化水平明显受到抑制（P<0.05）。因此，肝癌组织中miR-101的表达水平下降，miR-101过表达可抑制肝癌细胞的增殖。miR-101对肝癌细胞增殖的抑制作用可能与其对丝裂原活化蛋白激酶（MAPK）/ERK信号通路的抑制有关，而对MAPK/ERK的抑制可能是由miR-101对EZH2的靶向抑制介导的。
