〔Abstract〕 Objective To understand the bioinformatics of human ribosomal protein L22(RPL22) gene, prokaryotic expression and purification of human RPL22 recombinant protein, synthesis of human RPL22(labeled as agent M)in vitro, study the effects of agent M on proliferation, cycle and apoptosis of NSCLC A549 cells, and to analyze the effect of combination chemotherapy with M and cisplatin(DDP). Methods RPL22 bioinformatics was analyzed. RPL22 gene was cloned by PCR, the single chain oligonucleotide was designed and synthesized to obtain the target gene, and connect the pET-28α vector. The pET-28α-RPL22 recombinant plasmid was constructed, and the receptive cell DH5αof Escherichia coli was transformed. The expression of recombinant protein RPL22 was analyzed by SDS-PAGE and Western blot. The semi-inhibitory rate(IC50) of M and DDP in A549 cells and the effect of M on the proliferation of A549 cells were detected by CCK-8. The effect of agent M on apoptosis and cycle of A549 cells was detected by flow cytometry. qPCR and Western blot assays were used to detect M and DDP on phosphatidylinositol-3-kinase/protein kinase B(PI3K/AKT) and adenosine 5′-monophosphate activation of protein kinase/mammalian target of rapamycin(AMPK/mTOR) signaling pathway, respectively. Results RPL22 protein was an unstable and hydrophilic protein. The recombinant protein was dissolved and purified to obtain high concentration of recombinant protein. In A549 cells, the IC50of reagent M was 400 μg/L, and the IC50of DDP was 10 μmol/L, and the concentration of reagent M was inversely proportional to the cell activity. The inhibitory effect of reagent M(200 μg/L) combined with DDP(2.5 μmol/L) was similar to that of DDP(10 μmol/L) alone. Reagent M could induce G2cell cycle arrest and promote apoptosis, and might be involved in the regulation of PI3K/AKT and AMPK/mTOR pathways. Conclusion The recombinant RPL22 protein is cloned and expressed successfully.