Cheng Chen, Xiaohua Wang, Ning Jiang, Dan Zong, Chengyun Yao, Lijun Zhao, Li Yin, Shengfu Huang, Hong Ji, He Xia
Department of Radiotherapy, Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & The Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, 210009, China.
Department of Oncology, Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & The Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, 210009, China.
Cell Mol Biol (Noisy-le-grand). 2018 Oct 30;64(13):21-25.
To investigate the effects of microRNA-122 (miR-122) on the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) HONE-1 cells, and its correlation with the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. Human NPC cell line (HONE-1) was transfected with miR-122 inhibitor (anti-miR-122 group), negative controls (vector control group) via lipofectamines, and HONE-1 cell lines undergoing no transfection were selected (non-transfection group). The expression of miR-122, cell proliferation, apoptosis, and expressions of PI3K/AKT pathway and downstream target proteins in the three groups were determined using fluorescence quantitative polymerase chain reaction (qPCR), cell counting kit-8 (CCK8), immunofluorescence (IF) and Western blotting, respectively. The expression of miR-122 in the anti-miR-122 group was significantly lower than corresponding expressions in the non-transfection and vector control groups after 48h of transfection (p <0.05). The proliferation of cells in the anti-miR-122 group was significantly reduced with time after transfection (p <0.05). After 48h of transfection, the extent of apoptosis in the anti-miR-122 group (47.11 ± 1.95%) was significantly higher than that in normal control (7.37 ± 0.82%) and vector control group (8.54 ± 0.96%; p <0.05). There were no significant differences in the expressions of PI3K, AKT, mTOR protein, and the downstream signal proteins (p70S6K and 4E-BP1) in the three groups (p >0.05). However, the expressions of phosphorylated forms of these proteins were significantly lower in the anti-miR-122 group than in the non-transfection and vector control groups (p <0.05). IF results revealed that there were no significant differences in the fluorescence intensity value of PI3K and Akt among the three groups of patients (p>0.05). Inhibition of the expression of miR-122 in NPC suppresses the proliferation, and promotes their apoptosis through the PI3K/AKT signal transduction pathway.
探讨微小RNA-122（miR-122）对鼻咽癌（NPC）HONE-1细胞增殖和凋亡的影响及其与磷酸肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路的相关性。采用脂质体分别将miR-122抑制剂转染人NPC细胞系（HONE-1）（抗miR-122组）、阴性对照（载体对照组），选取未转染的HONE-1细胞系（未转染组）。分别采用荧光定量聚合酶链反应（qPCR）、细胞计数试剂盒-8（CCK8）、免疫荧光（IF）和蛋白质免疫印迹法检测三组中miR-122的表达、细胞增殖、凋亡以及PI3K/AKT通路及其下游靶蛋白的表达。转染48h后，抗miR-122组中miR-122的表达显著低于未转染组和载体对照组（p<0.05）。转染后抗miR-122组细胞增殖随时间显著降低（p<0.05）。转染48h后，抗miR-122组的凋亡率（47.11±1.95%）显著高于正常对照组（7.37±0.82%）和载体对照组（8.54±0.96%；p<0.05）。三组中PI3K、Akt、mTOR蛋白及下游信号蛋白（p70S6K和4E-BP1）的表达无显著差异（p>0.05）。然而，抗miR-122组中这些蛋白的磷酸化形式表达显著低于未转染组和载体对照组（p<0.05）。IF结果显示，三组患者PI3K和Akt的荧光强度值无显著差异（p>0.05）。抑制NPC中miR-122的表达可抑制其增殖，并通过PI3K/AKT信号转导通路促进其凋亡。
