[Inhibitory effects of targeting protein kinase B1 and cyclooxygenase-2 shRNA upon human gastric adenocarcinoma cell growth].

OBJECTIVE

To construct a short hairpin RNA (shRNA) adenovirus vector targeting Akt1 (protein kinase B1, PKB1/Akt1) and cyclooxygenase-2 (COX-2) and study its effects on the growth of SGC-7901 human gastric adenocarcinoma cell.

METHODS

Adenovirus pGSadeno-Akt1 + COX-2 (rAd5-A + C) vector was constructed and transfected into SGC-7901 cell. The proliferative activity of tumor cell was evaluated by MTT assay and flow cytometry in vitro. rAd5-HK and rAd5-A + C were injected into the established subcutaneous SGC-7901 gastric adenocarcinoma in nude mice. During the observation period of 21 days, tumor volume was measured every 3 days to further observe the anti-tumor effects of rAd5-A + C on SGC-7901 cell and cell situ apoptosis was detected by TUNEL assay.

RESULTS

After transfection of constructed adenovirus vector rAd5-A + C into SGC-7901 cell, cell proliferative activity in rAd5-A + C treatment group was significantly suppressed, and cell cycle indicated that control group SGC-7901 and no-load group rAd5-A + C cells in G0/G1, S and G2/M phases accounted for the total number of cells 49.8%, 35.2%, 15.0% and 50.8%, 36.5%, 12.7% respectively. While the treatment group rAd5-A + C in G0/ G1, S and G2/M phases accounted for the total number of cells 68.1%, 21.8% and 10.1% respectively. The tumor volume in treatment group was lower than that of control and no-load groups and the difference had statistical significance (F = 3.679, P = 0.043) and rAd5-A + C could induce the apoptosis of tumor cell.

CONCLUSION

Adenovirus-mediated Akt1 and COX-2 shRNA can inhibit the growth of SGC-7901 human gastric adenocarcinoma cell. It may provide a new strategy for gastric cancer gene therapy.