|
|
|
| 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. |
|
|
|
