Preliminary Investigation Of The Effect Of NYD-Tr And NYD-Ch Genes Of Deltamethrin Resistant Culex Pipiens Pallens, To Human Tumor Cells JAR And HepG2

[Object] order to investigate the effect of NYD-Tr and NYD-Ch genes of deltamethrin resistance associated Cx.pipiens pallens, to human choriocarcinoma cells and Hepatocarcinoma cells in vitro.[Materials and methods 1 Two pairs of primers were designed according to NYD-Tr and NYD-Ch genes released by our laboratory in GenBank (AF034060 and AF468495). The ORF sequences were cloned into eukaryon expression plasmid pEGFP-C3 which has C-Terminal Fluorescent Protein (EGFP) as reporter gene. After the recombined plasmids were got, JAR cell line(human choriocarcinoma cell line) was transfected by Fugene 6 and HepG2 cell line (human hepatocellular carcinoma cell line grown in a laboratory environment) was transfected by Effectene Reagent. Fluorescences were observed by Fluorescence inversion microscope. The configurations of cells and expression of Green Fluorescence were taken pictures by Laser confocal microscopy. G418 was used to screen the positive cells. MTT and electron microscope were used to study the positive cells.[Results ] By sequencing and enzyme cutting, the genes of NYD-Tr and NYD-Ch we obtained were identical with the released genes in GenBank and expressed in the same ORF with EGFP. The recombinant plasmids pEGFP-C3/ NYD-Tr and pEGFP-C3/ NYD-Ch were transfected successfully. The transfection efficiency of Fugene 6 was higher than that of Effectene Reagent. Pictures taken by Laser confocal microscopyshowed that the cells co-expressed EGFP and NYD-Tr/ NYD-Ch were affected in morphology. The results of MTT displayed that the growth rates of JAR cells transfected with pEGFP-C3/ NYD-Tr and pEGFP-C3/ NYD-Ch were lower 39.51% and 45.88% compared to the cells transfected with pEGFP-C3, respectively, and the growth rates of HepG2 cells transfected with pEGFP-C3/ NYD-Tr and pEGFP-C3/ NYD-Ch were lower 14.39% and 63.13% respectively. The JAR cells transfected with pEGFP-C3/ NYD-Tr and pEGFP-C3/ NYD-Ch were damnified severely by electron microscope.[ Conclusion ] NYD-Tr and NYD-Ch, the deltamethrin resistance associated genes of Cx. pipiens pallcns, can restrain and scathe the human tumor cells JAR and HepG2 in vitro. To confirmation the effect and to explore the mechanism needs to go deep into the research.