Inhibiting effects on HeLa cell growth by HER-2 gene targeting

A plasmid (promoter pPNT 1305) for HER-2 gene targeting was constructed and introduced into HeLa cells by electroporation. After positive selection by G418 and negative selection by Gancyclovir, the targeted cell line (HER-2( -1) cells) containing only one copy of HER-2 allies was selected. Northern blot assay showed that the mRNA expression of HER-2(-1) was significantly l ess than that of its parental cells. The results of Western blot also showe d that HER-2 protein of HER-2(-1) cells was obviously less than that of HeL a cells. The data of FACS assay demonstrated that after one of the two HER- 2 copies in HeLa cells was knocked out, the G1 phase percentage of HER-2(-1 ) cells was more and the M plus G2 phase percentage of HER-2(-1) L cells wa s less than those of HeLa cells respectively. These results show that gene targeting of HER-2 has exhibited the inhibiting effects on the proliferatio nal capability of HeLa cells.