Suppression of tumorigenicity of HepG2.2.15 cells by N-ras antisense RNA transfected with AFP-enhancing four-element complex
Objective To observe the inhibitory effects of N-ras antisense RNA delivered by AFP-enhancing four-element complex on tumorigenicity of HBV-transgenic hepatocellular carcinoma cell line HepG 2.2.15 both in vitro and in vivo . Methods AFP-enhancing four-element complex,containing N-ras antisense RNA, was constructed and transfected into HepG2.2.15 cells. Seventy-two hours later, cells were collected and assayed for expression of N-ras protein, cell cycle distribution and apoptosis rate by flow cytometry. In addition, HepG 2.2.15 /as-ras,stably expressing N-ras antisense RNA, was established. The growth curve of two cell lines was delineated. BALB/c nude mice were implanted with HepG2.2.15 or HepG2.2.15/as-ras cells. The tumor-forming rate for two groups were calculated. BALB/c nude mice bearing with HepG2.2.15 cells was injected with AFP-enhancing four-element complex including N-ras antisense RNA and the volumes of tumor were measured. Results After transfection by N-ras antisense RNA four-element complex, the expression of N-ras protein of HepG2.2.15 cells was greatly decreased; cell cycle was trapped at G 0/G 1 phase; apoptosis rate was enhanced. Compared with HepG2.2.15 cells, the tumor-forming rate of HepG2.2.15/as-ras cells was lower. When HepG 2.2.15 cells were implanted into nude mice, injection of N-ras antisense RNA four-element complex could significantly decrease the volume of tumor tissues. Conclusion AFP-enhancing four-element complex including N-ras antisense RNA inhibited the expression of N-ras protein and effectively impaired the tumorigenicity of HepG2.2.15 cells both in vitro and in vivo .