The eastern Himalayas are located near the southern entrance through which early modern humans expanded into East Asia. The genetic structure in this region is therefore of great importance in the study of East Asian origins. However, few genetic studies have been performed on the Sino-Tibetan populations (Luoba and Deng) in this region. Here, we analyzed the Y-chromosome diversity of the two populations. The Luoba possessed haplogroups D, N, O, J, Q, and R, indicating gene flow from Tibetans, as well as the western and northern Eurasians. The Deng exhibited haplogroups O, D, N, and C, similar to most Sino-Tibetan populations in the east. Short tandem repeat (STR) diversity within the dominant haplogroup O3 in Sino-Tibetan populations showed that the Luoba are genetically close to Tibetans and the Deng are close to the Qiang. The Qiang had the greatest diversity of Sino-Tibetan populations, supporting the view of this population being the oldest in the family. The lowest diversity occurred in the eastern Himalayas, suggesting that this area was an endpoint for the expansion of Sino-Tibetan people. Thus, we have shown that populations with haplogroup O3 moved into the eastern Himalayas through at least two routes.