For the purpose of understanding the self-assembly formation mechanism of DNA base molecules, guanine, adenine, cytosine, and thymine molecules were deposited on Cu(111) surfaces, and were observed using a low-temperature (approximate to 80 K) scanning tunneling microscope (STM). Single-molecular-scale STM images revealed that guanine, adenine, and cytosine molecules can form ordered one- and/or two-dimensional unique structures, but thymine molecules, however, randomly aggregate into small clusters. Semiempirical molecular orbital (MO) calculation indicates that there exists predominantly stable dimer structures for the former three molecules, while such phenomena cannot be observed among the possible thymine dimer and even trimer structures. Based on experimental and theoretical results, we have concluded that specific hydrogen-bonded nucleus formation is a decisive process in the two-dimensional self-assembly formation of DNA base molecules on Cu(111) surfaces.