In order to determine the combined effects of migration and gene flow on evolution of insecticide resistance in the mosquito Culex pipiens, four samples were collected in China, among which two were collected along the railway from Beijing to Guangzhou. Bioassay data showed that the resistance levels of the four populations to dichlorvos were high and to parathion moderate as compared with the susceptible strain and there was no significant difference among the four populations to the same organophosphate (OP) insecticide. Starch electrophoresis was done to identify the frequency of known overproduced esterases and to analyze genetic diversity among various populations by electrophoretic polymorphism of five presumably neutral loci. The results indicated that the gene flow between populations existed and the number of effective migrants (Nm) was related to collection geography (Nm from 1.67 to 40.07). In contrast with lower genetic differentiation between two nearby populations (between GZ and ZS, ZZ and SQ) and higher genetic differentiation between two distant populations (between GZ and ZZ), there was a significant and inconsistent difference in the distribution of resistance alleles, A2-B2 when explained only with active migration. This divergent situation could be straightened out when considering passive migration (such as railway transport) which increased the spread of A2-B2 along the railway, i.e., in GZ and ZZ. The resistance alleles, A2-B2, dispersing to around areas by active migration suffered from the limitation of gene flow and the speed of invasion.