Effect of lentivirus targeted to bring Akt1 shRNA on apoptosis and proliferation of gastric carcinoma SGC-7901 Cells
TAN Wei YU Honggang LUO Hesheng ZHOU Wei▲
Department of Gastroenterology, Renmin Hospital of Wuhan University Hubei Key Laboratory of Digestive System Disease, Hubei Province, Wuhan 430060, China
Abstract：Objective To investigate the apoptosis and proliferation effect of lentiviral vectors shRNA for Akt1 to human gastric carcinoma SGC-7901 cells. Methods The lentiviral vectors Akt1-shRNA were constructed, lentivirus Akt1-shRNA was transfected into human gastric carcinoma SGC-7901 cells, the SGC-7901 cell strained with Akt1 gene silencing perpetually (SGC-LV-shCON) was as conrol group cell and amplified (SGC LV-shAkt1) was as experiment group cell. Effect of Akt1 on SGC-7901 cells were measured by RT-PCR, Western-blot, flow cytometry of annexin V-FITC/PI, tumor growth curve. Results Akt1 RNAi lentiviral vectors LV-shAkt1 was established successfully, virus titer was 5×108 TU/mL. After SGC-7901 cell transfection, the expression of Akt1 mRNA in experiment group (SGC- LV-shAkt1) was weaker than that in the control group (SGC-7901, SGC- LV-shCON), and Akt1, p-Akt and bcl-2 protein expressions in experiment group was inhabited (P < 0.01), (38.7±3.5)% cell in experiment group happened apotosis (P < 0.05). The cell growth curve of SGC-LV-shAkt1 cell was gentle, compared with SGC-7901 cell or SGC- LV-shCON cell, the growth was lower. Conclusion The lentiviral vectors mediated shRNA for Akt1 can significantly inhibit the expression of Akt1 gene in vitro and markedly inhibit SGC-7901 cell growth and induce the apoptosis of gastric carcinoma cell.The mechanism may be down-regulation of PI3K/Akt signal pathway and anti-apoptosis protein bcl-2 expression.
[1] Jin SY，Lee HS，Kim EK，et al. Reactive oxygen species and PI3K/Akt signaling in cancer [J]. Free Radic Biol Med，2014，75（Suppl 1）：S34-S35.
[2] Hay M. Interplay between FOXO，TOR，and Akt [J]. Biochim Biophys Acta，2011，1813（11）：1965-1970.
[3] Hernandez-Aya LF，Gonzalez-Angulo AM. Targeting the phosphatidylinositol 3-kinase signaling pathway in breast cancer [J]. Oncologist，2011，16（4）：404-414.
[4] Sarkies P，Miska EA. RNAi pathways in the recognition of foreign RNA：antiviral responses and host-parasite interactions in nematodes [J]. Biochem Soc Trans，2013，41（4）：876-880.
[5] Saito T，Kurokawa Y，Takiguchi S，et al. Current status of function-preserving surgery for gastric cancer [J]. World J Gastroenterol，2014，20（46）：17297-17304.
[6] Zhuo B，Li Y，Li Z，et al PI3K/Akt signaling mediated Hexokinase-2 expression inhibits cell apoptosis and promotes tumor growth in pediatric osteosarcoma [J]. Biochem Biophys Res Commun，2015，464（2）：401-406.
[7] Zhou W，Fu XQ，Zhang LL，et al. The AKT1/NF-kappaB/Notch1/PTEN axis has an important role in chemoresistance of gastric cancer cells [J]. Cell Death Dis，2013，4（10）：e847.
[8] Agarwal E，Brattain MG，Chowdhury S. Cell survival and metastasis regulation by Akt signaling in colorectal cancer [J]. Cell Signal，2013，25（8）：1711-1719.
[9] Davies MA，Koul D，Dhesi H，et al. Regulation of Akt/PKB activity，cellular growth，and apoptosis in prostate carcinoma cells by MMAC/PTEN [J]. Cancer Res，1999，59（11）：2551-2556.
[10] Dykxhoorn DM. RNA interference as an anticancer therapy：a patent perspective [J]. Expert Opin Ther Pat，2009， 19（4）：475-491.
[11] Shen T，Liu K，Miao D，et al. Lentivirus-mediated RNA interference against Japanese encephalitis virus infection in vitro and in vivo[J]. Antiviral Res，2014，108：56-64.
[12] 朴莲淑，王福光，于庆功，等.稳定抑制PAK2蛋白表达的HUH-7细胞株的建立[J].中国医药导报，2015，12（27）：14-17.
[13] 段俊鹏，宋建，范现英，等.CXCR4介导胃癌细胞的抗凋亡机制的研究进展[J].中国现代医生，2015，53（10）：156-160.
[14] Xie X，Tang B，Zhou J，et al. Inhibition of the PI3K/Akt pathway increases the chemosensitivity of gastric cancer to vincristine [J]. Oncol Rep，2013，30（2）：773-782.
[15] 吴松，虞桂平，薛涛，等.PI3K-mTOR信号通路抑制剂抗肿瘤研究新进展[J].中国医药导报，2015，12（1）：161-164，168.
2017, Vol. 14 
京公网安备 11010502046598号