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Int J Clin Exp Pathol 2013;6(10):2071-2081

Original Article
Small interfering RNA survivin and GRIM-19 co-expression salmonella plasmid inhibited
the growth of laryngeal cancer cells in vitro and in vivo

Lian-Ji Wen, Li-Fang Gao, Chun-Shun Jin, He-Jia Zhang, Kun Ji, Jing-Pu Yang, Xue-Jian Zhao, Ming-Ji Wen, Guo-Fang Guan

Department of Otorhinolaryngology, Second Hospital of Jilin University, Changchun 130041, China; WeiKang Hospital of Shenyang, Shenyang
110000, China; Department of Ultrasonography, China-Japan Union Hospital of Jilin University, Changchun 130033, China; Department of
Pathophysiology, Medicinal University of Shenyang, Shenyang 110034, China; Department of Pathophysiology, Basic School of Medicine, Jilin
University, Changchun 130021, China; Department of Pharmacy, Hospital of Changchun City, Changchun 130051, China

Received August 4, 2013; Accepted August 27, 2013; Epub September 15, 2013; Published October 1, 2013

Abstract: Objective: To investigate the inhibitory effect of plasmid-based survivin-specific short hairpin RNA and GRIM-19 on the growth of
Hep-2 laryngeal cancer cells. Methods: The plasmid expressing survivin-specific short hairpin RNA (shRNA) and GRIM-19 (p-siRNA
survivin/GRIM-19) was prepared and transfected into Hep-2 cells with Lipofectamine 2000. The mRNA and protein expression of surviving and
GRIM-19 were measured with RT-PCR and western blot assay, respectively. MTT assay was employed to detect the proliferation of Hep-2 cells,
and flow cytometry and AO/EB assay were done to determine the apoptosis of Hep-2 cells. Results: In the p-siRNA survivin/GRIM-19, the mRNA
and protein expression of survivin was markedly reduced by 54.4% and 42.2%, and the reduction in protein expression of surviving was more
obvious than that in the p-siRNA survivin group (37%) (P<0.05). The protein expression of GRIM-19 was markedly enhanced when compared
with the control group (P<0.01). MTT assay revealed the proliferation of Hep-2 cells undergoing transfection with p-siRNA survivin/GRIM-19 was
markedly inhibited, and the inhibition rate was as high as 79%, which was higher than that in the psi-survivin group (45%) and p-GRIM-19
group (35%). AO/EB assay and flow cytometry indicated that the apoptotic cells in the p-siRNA survivin/GRIM-19 group were dramatically
increased as compared to the psi-survivin group and p-GRIM-19 group. Conclusion: The p-siRNA survivin/GRIM-19 has marked decrease in
survivin expression and dramatic increase in GRIM-19 expression. Moreover, silencing of survivin and over-expression of GRIM-19 can
significantly inhibit the growth and induce the apoptosis of Hep-2 in vitro and in vivo. (IJCEP1308010).

Keywords: Co-expression plasmid, gene silencing, survivin, laryngeal cancer

Address correspondence to: Guo-Fang Guan, Department of Otorhinolaryngology, Second Hospital of Jilin University, Changchun 130041,
China. E-mail: guofangguan82@sina.com