miR-222 is necessary for exercise-induced cardiac growth and protects against pathological cardiac remodeling.. Rattus norvegicus

miR-222 overexpression leads to promotion of proliferation and hypertrophy and inhibition of apoptosis in in primary neonatal rat ventricular cardiomyocytes (NRVMs). Isolated primary neonatal rat ventricular cardiomyocytes were plated in 6 cm BD Primaria tissue culture dishes. Transfection of microRNA precursors or scramble control (0.4 μM) was carried out using Lipofectamine RNAiMAX (Invitrogen) as recommended by the manufacturer. Forty-eight hours after transfection, RNAs from cultured cells and tissues were isolated with Tryzol (Invitrogen) following the manufactures’ manuals. Total RNA was harvested and submitted to the Dana-Farber Cancer Institute Molecular Diagnostics Laboratory for assay. These results revealed miR-222 regulated gene expression in primary neonatal rat ventricular cardiomyocytes. Overall design: Gene expression in NRVM cells tranfected with control scramble precursor (4 samples) or miR-222 precursor (4 smaples ) was evaluated using Affymetrix Rat Genome 230 v2.0.

miR-222过表达可促进原代新生大鼠心室心肌细胞（primary neonatal rat ventricular cardiomyocytes, NRVMs）的增殖与肥大，并抑制其凋亡。将分离得到的原代新生大鼠心室心肌细胞接种于6 cm BD Primaria组织培养皿中。按照制造商推荐的操作流程，使用Lipofectamine RNAiMAX（Invitrogen）转染miRNA前体或阴性对照scramble control（浓度为0.4 μM）。转染48小时后，按照制造商提供的操作手册，使用Tryzol（Invitrogen）提取培养细胞与组织中的总RNA。收集总RNA并送至丹娜-法伯癌症研究所（Dana-Farber Cancer Institute）分子诊断实验室进行检测分析。上述结果表明，miR-222可调控原代新生大鼠心室心肌细胞的基因表达。总体实验设计：采用Affymetrix大鼠基因组230 v2.0芯片（Affymetrix Rat Genome 230 v2.0），对转染阴性对照scramble前体的NRVM细胞（4个样本）与转染miR-222前体的NRVM细胞（4个样本）的基因表达水平进行评估。