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. 2006 May 14;12(18):2901-7.
doi: 10.3748/wjg.v12.i18.2901.

Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity

Hai-Tao Guan  1 , Xing-Huan Xue, Zhi-Jun Dai, Xi-Jing Wang, Ang Li, Zhao-Yin Qin
Affiliations

Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity

Hai-Tao Guan et al. World J Gastroenterol. .

Abstract

Aim: To investigate the inhibitory effect of small interfering RNA (siRNA) on the expression of survivin in pancreatic cancer cell line PC-2 and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity.

Methods: A siRNA plasmid expression vector against survivin was constructed and transfected into PC-2 cells with Lipofectamine 2000. The down regulation of survivin expression was detected by semi-quantitative RT-PCR and immunohistochemical SP method and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity was detected by flow cytometry.

Results: The sequence-specific siRNA efficiently and specifically down-regulated the expression of survivin at both mRNA and protein levels. The expression inhibition ratio was 81.25% at mRNA level detected by semi-quantitative RT-PCR and 74.24% at protein level detected by immunohistochemical method. Forty-eight hours after transfection,apoptosis was induced in 7.03% cells by siRNA and in 14.58% cells by siRNA combined with radiation.

Conclusion: The siRNA plasmid expression vector against survivin can inhibit the expression of survivin in PC-2 cells efficiently and specifically. Inhibiting the expression of survivin can induce apoptosis of PC-2 cells and enhance its radiosensitivity significantly. RNAi against survivin is of potential value in gene therapy of pancreatic cancer.

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Figures

Figure 1
Figure 1
Reports of the sequencing of plasmid Pgenesil-sur(+) (A) and plasmid Pgenesil-sur (-) (B).
Figure 2
Figure 2
Expression of EGFP in PC-2 cells 24 h after trancfection of plasmid Pgenesil-sur(+) (A) and plasmid Pgenesil-sur(-) (B) (original magnification×200).
Figure 3
Figure 3
Down-regulation of survivin mRNA expression detected by semi-quantitive RT-PCR. 1: The blank control group; 2: The negative control group; M: Marker I; 3: The positive experiment group.
Figure 4
Figure 4
Down-regulation of survivin expression detected by immunohisto- chemical SP method in the blank control group (A), negative control group (B), and positive experiment group (C) (coloration with DAB, original magnification × 400)
Figure 5
Figure 5
Apoptosis of PC-2 cells detected in 6 groups by flow cytometry (stained with PI)
See this image and copyright information in PMC

References

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