Purpose　Non-small　cell　lung　cancer　(NSCLC)　is　the　most　common　type　of　lung　cancer　and　ranked　top　in　terms　of　incidence　and　mortality　in　men　and　women.　Recently,　improvements　in　treatment　approaches　for　NSCLC　have　reported,　but　still,　there　is　a　need　to　devise　innovative　treatment　strategies,　especially　to　manage　the　advanced　and　metastatic　stage　of　NSCLC.　Aloperine　(ALO),　an　herbal　alkaloid,　has　exerted　anti-cancer　effects　in　many　cancers.　However,　the　use　of　any　chemotherapeutic　agents　is　dose　limited　due　to　possible　adverse　effects　and　drug-resistance　issues.　Therefore,　a　combination　of　chemotherapy　with　viral-based　targeted　gene　therapy　may　provide　a　novel　treatment　strategy　for　NSCLC.　Methods/results　In　this　study,　the　results　of　the　MTT　and　flow　cytometry-based　assays　showed　that　Aloperine-Adbic　(adenoviral　vector　expressing　p14(ARF)/p53)　combined　treatment　on　NSCLC　cells　synergistically　produced　anti-proliferative　effects,　induced　apoptosis,　and　arrested　cell　cycle　at　the　G1　phase.　Furthermore,　the　expression　analysis　suggested　that　the　p53/p21　pathway　might　contribute　to　achieving　aforesaid　cytotoxic　effects.　The　ALO-Adbic　combined　treatment　prolonged　the　percent　survival　of　NSCLC　xenograft　models.　Conclusion　In　conclusion,　ALO-Adbic　combination　can　produce　synergistic　anti-cancer　effects　at　low　doses,　and　may　offer　a　more　effective　and　less　toxic　new　treatment　strategy　for　NSCLC.