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Reverse Transcription Quantitative PCR (RT-qPCR) Following total RNA extraction using TRIzol reagent (Invitrogen), cDNA was synthesized using Mir-X miRNA First-Strand Synthesis Kit (Takara Holdings Inc., Kyoto, Japan) with extracted total RNA (2 μg). The miRNA expression was measured by qPCR with SYBR Advantage qPCR Premix (Takara) on CFX133a Real-Time PCR Detection System (Bio-Rad Laboratories, Hercules, CA, USA). Relative expression was determined with a U6 control through the 2 −ΔΔCt method. The primers used are depicted in Table 1. Table 1 Primer Sequences Cell Culture, Transfection and Infection Human lung cancer cell lines, including H522, H460, SK-MES-1, and A549, and a human bronchial epithelial cell line BEAS-2B, were from the Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). Cell lines H522, H460, and A549 were cultivated in RPMI-1640 medium (Gibco; Thermo Fisher Scientific), while SK-MES-1 cells were in minimal medium (Gib

United statesSwitzerland generalUnited kingdomEteläuomen läiSanta claraLonza cloneticsLaboratory animal co ltdCommittee of people hospital zhengzhou universityChinese academy of sciences shanghaiBiotech co ltdPeople hospital of zhengzhou universityInvitrogen incCo ltdOlympus optical co ltdInstitute of biochemistryStrand synthesis kit takara holdings inc